GenBank-Updates@genbank.bio.net (04/12/91)
LOCUS ECOTGP 7539 bp ds-DNA BCT 12-APR-1991 DEFINITION E.coli tryptophan operon: entire DNA sequence. ACCESSION J01714 M12471 M12472 M25593 M59208 KEYWORDS anthranilate isomerase; anthranilate synthetase; attenuator; glutamine amidotransferase; isomerase; leader peptide; phosphoribosyl anthranilate synthetase; synthetase; transferase; trp operon; trpA gene; trpB gene; trpC gene; trpD gene; trpE gene; tryptophan synthetase. SOURCE Escherichia coli RNA and DNA. ORGANISM Escherichia coli Prokaryota; Bacteria; Gracilicutes; Scotobacteria; Facultatively anaerobic rods; Enterobacteriaceae. REFERENCE 1 (bases 5917 to 6133) AUTHORS Platt,T. and Yanofsky,C. TITLE An intercistronic region and ribosome-binding site in bacterial messenger RNA JOURNAL Proc. Natl. Acad. Sci. U.S.A. 72, 2399-2403 (1975) STANDARD full staff_review REFERENCE 2 (bases 84 to 141) AUTHORS Bennett,G.N., Schweingruber,M.E., Brown,K.D., Squires,C. and Yanofsky,C. TITLE Nucleotide sequence of region preceding trp mRNA initiation site and its role in promoter and operator function JOURNAL Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976) STANDARD full staff_review REFERENCE 3 (bases 117 to 310) AUTHORS Squires,C., Lee,F., Bertrand,K., Squires,C.L., Bronson,M.J. and Yanofsky,C. TITLE Nucleotide sequence of the 5' end of tryptophan messenger RNA of Escherichia coli JOURNAL J. Mol. Biol. 103, 351-381 (1976) STANDARD full staff_review REFERENCE 4 (bases 230 to 272) AUTHORS Bertrand,K., Korn,L.J., Lee,F. and Yanofsky,C. TITLE The attenuator of the tryptophan operon of Escherichia coli: heterogeneous 3'-OH termini in vivo and deletion mapping of functions JOURNAL J. Mol. Biol. 117, 227-247 (1977) STANDARD full staff_review REFERENCE 5 (bases 230 to 272) AUTHORS Stauffer,G.V., Zurawski,G. and Yanofsky,C. TITLE Single base-pair alterations in the Escherichia coli trp operon leader region that relieve transcription termination at the trp attenuator JOURNAL Proc. Natl. Acad. Sci. U.S.A. 75, 4833-4837 (1978) STANDARD full staff_review REFERENCE 6 (bases 6707 to 6863) AUTHORS Wu,A.M. and Platt,T. TITLE Transcription termination: nucleotide sequence at 3' end of tryptophan operon in Escherichia coli JOURNAL Proc. Natl. Acad. Sci. U.S.A. 75, 5442-5446 (1978) STANDARD full staff_review REFERENCE 7 (bases 0 to 0) AUTHORS Bennett,G.N., Schweingruber,M.E., Brown,K.D., Squires,C. and Yanofsky,C. TITLE Nucleotide sequence of the promoter-operator region of the tryptophan operon of Escherichia coli JOURNAL J. Mol. Biol. 121, 113-137 (1978) STANDARD full staff_review REFERENCE 8 (bases 36 to 136) AUTHORS Brown,K.D., Bennet,G.N., Lee,F., Schweingruber,M.E. and Yanofsky,C. TITLE RNA polymerase interaction at the promoter-operator region of the tryptophan operon of Escherichia coli and Salmonella typhimurium JOURNAL J. Mol. Biol. 121, 153-177 (1978) STANDARD simple staff_entry REFERENCE 9 (bases 2351 to 2503) AUTHORS Miozzari,G.F. and Yanofsky,C. TITLE Gene fusion during the evolution of the tryptophan operon in enterobacteriaceae JOURNAL Nature 277, 486-489 (1979) STANDARD full staff_review REFERENCE 10 (bases 5932 to 6809) AUTHORS Nichols,B.P. and Yanofsky,C. TITLE Nucleotide sequences of trpA of Salmonella typhimurium JOURNAL Proc. Natl. Acad. Sci. U.S.A. 76, 5244-5248 (1979) STANDARD full staff_review REFERENCE 11 (bases 117 to 256) AUTHORS Oxender,D.L., Zurawski,G. and Yanofsky,C. TITLE Attenuation in the Escherichia coli tryptophan operon: role of RNA secondary structure involving the tryptophan codon region JOURNAL Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979) STANDARD full staff_review REFERENCE 12 (bases 6707 to 7335) AUTHORS Wu,A.M., Chapman,A.B., Platt,T., Guarente,L.P. and Beckwith,J. TITLE Deletions of distal sequence affect termination of transcription at the end of the tryptophan operon in E. coli JOURNAL Cell 19, 829-836 (1980) STANDARD full staff_review REFERENCE 13 (bases 230 to 296) AUTHORS Farnham,P.J. and Platt,T. TITLE A model for transcription termination suggested by studies on the trp attenuator in vitro using base analogs JOURNAL Cell 20, 739-748 (1980) STANDARD full staff_review REFERENCE 14 (bases 4810 to 6003) AUTHORS Crawford,I.P., Nichols,B.P. and Yanofsky,C. TITLE Nucleotide sequence of the trpB gene in Escherichia coli and Salmonella typhimurium JOURNAL J. Mol. Biol. 142, 489-502 (1980) STANDARD full staff_review REFERENCE 15 (bases 1761 to 2443) AUTHORS Nichols,B.P., Miozzari,G.F., van Cleemput,M., Bennett,G.N. and Yanofsky,C. TITLE Nucleotide sequences of the trpG regions of Escherichia coli, Shigella dysenteriae, Salmonella typhimurium and Serratia marcescens JOURNAL J. Mol. Biol. 142, 503-517 (1980) STANDARD full staff_review REFERENCE 16 (bases 3422 to 4824) AUTHORS Christie,G.E. and Platt,T. TITLE Gene structure in the tryptophan operon of Escherichia coli: nucleotide sequence of trpC and the flanking intercistronic regions JOURNAL J. Mol. Biol. 142, 519-530 (1980) STANDARD full staff_review REFERENCE 17 (bases 5932 to 6809) AUTHORS Schneider,W.P., Nichols,B.P. and Yanofsky,C. TITLE Procedure for production of hybrid genes and proteins and its use in assessing significance of amino acid differences in homologous tryptophan synthetase alpha polypeptides JOURNAL Proc. Natl. Acad. Sci. U.S.A. 78, 2169-2173 (1981) STANDARD full staff_review REFERENCE 18 (bases 6807 to 6856; 7057 to 7119) AUTHORS Wu,A.M., Christie,G.E. and Platt,T. TITLE Tandem termination sites in the tryptophan operon of Escherichia coli JOURNAL Proc. Natl. Acad. Sci. U.S.A. 78, 2913-2917 (1981) STANDARD full staff_review REFERENCE 19 (bases 279 to 1843) AUTHORS Nichols,B.P., van Cleemput,M. and Yanofsky,C. TITLE Nucleotide sequence of Escherichia coli trpE: anthranilate synthetase component I contains no tryptophan residues JOURNAL J. Mol. Biol. 146, 45-54 (1981) STANDARD full staff_review REFERENCE 20 (sites) AUTHORS Yanofsky,C., Platt,T., Crawford,I.P., Nichols,B.P., Christie,G.E., Horowitz,H., van Cleemput,M. and Wu,A.M. TITLE The complete nucleotide sequence of the tryptophan operon of Escherichia coli JOURNAL Nucleic Acids Res. 9, 6647-6668 (1981) STANDARD full staff_review REFERENCE 21 (bases 2504 to 3436) AUTHORS Horowitz,H., Christie,G.E. and Platt,T. TITLE Nucleotide sequence of the trpD gene, encoding anthranilate synthetase component II of Escherichia coli JOURNAL J. Mol. Biol. 156, 245-256 (1982) STANDARD full staff_review REFERENCE 22 (bases 57 to 137) AUTHORS Windass,J.D., Newton,C.R., De Maeyer-Guignard,J., Moore,V.E., Markham,A.F. and Edge,M.D. TITLE The construction of a synthetic Escherichia coli trp promoter and its use in the expression of a synthetic interferon gene JOURNAL Nucleic Acids Res. 10, 6639-6657 (1982) STANDARD full staff_review REFERENCE 23 (sites) AUTHORS Kolter,R. and Yanofsky,C. TITLE Genetic analysis of the tryptophan operon regulatory region using site-directed mutagenesis JOURNAL J. Mol. Biol. 175, 299-312 (1984) STANDARD full staff_review REFERENCE 24 (bases 1 to 350) AUTHORS Kane,J.F., Balaban,S.M. and Bogosian,G. TITLE Commercial production of bovine somatotropin in Escherichia coli JOURNAL (in) Sikes,C.S. and Wheeler,A.P. (Eds.); Surface reactive peptides and polymers. Discovery and commercialization.: In press, American Chemical Society, Washington, D.C. (1990) STANDARD simple staff_entry COMMENT [Nucleic Acids Res. 9, 6647-6668 (1981)] review; bases 77 to 6809; compiled. [J. Mol. Biol. 175, 299-312 (1984)] sites; mutational analysis of the regulatory region. The tryptophan operon of E.coli consists of a repressor(trpR), a promoter(trpP), an operator(trpO), an attenuator which is part of a leader peptide region(trpL) and five structural genes: trpE(anthranilate synthetase), trpD(glutamine amido transferase and anthranilate 5-phosphoribosylpyrophosphate phosphoribosyl- transferase), trpC(phosphoribosyl anthranilate isomerase-indole glycerol phosphate synthetase), trpB(tryptophan synthetase beta) and trpA(tryptophan synthetase alpha). The promoter region covers approximately 40 bases upstream from the mRNA initiation site(75-116); the operator approximately 20 bases upstream with two-fold axes of symmetry around 104-105 and 109-110([Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976)],[J. Mol. Biol. 121, 113-137 (1978)],[J. Mol. Biol. 156, 245-256 (1982)]). The attenuator region is the first 140 nucleotides(117-256) of the mRNA leader, a G-C rich region with a two-fold axis of symmetry around base 240 and an A-T rich region with its axis about bases 259-260; it provides a second site for control of transcription ([J. Mol. Biol. 117, 227-247 (1977)], [Proc. Natl. Acad. Sci. U.S.A. 75, 4833-4837 (1978)],[Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979)],[Cell 20, 739-748 (1980)]). Two mRNA termination regions are reported: trpT (bases 6807-6856) and trpT' (bases 7057-7119), the first of which bears some similarity to the attenuator region ([Proc. Natl. Acad. Sci. U.S.A. 78, 2913-2917 (1981)]). A chi site for recombination is localized between bases 2492 and 2501 and the trp-P2 promoter is located between bases 3240 and 3280 ([J. Mol. Biol. 156, 245-256 (1982)]). The trpE gene is unusual in that it codes for no tryptophan residues([J. Mol. Biol. 146, 45-54 (1981)]). The two enzymatic functions coded by trpG and trpD genes in S.marcescens are coded by the single trpD gene in E.coli and other enterobacteriaceae. This appears to have occurred via base changes at sites 2420 and 2438. The intercistronic regions for the structural genes show little superfluity: the trpE-trpD and trpB-trpA boundaries consist of 'tgatg'; the trpD-trpC boundary is 'taaatgatg' and the trpC-trpB boundary is 'taaggaaaggaacaatg'. All the cistrons show a high degree of homology with their correlates among the enterobacteriaceae. Sequence discrepancies in early work([J. Mol. Biol. 103, 351-381 (1976)]) are corrected in later work from the same laboratory([Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979)], [Nucleic Acids Res. 9, 6647-6668 (1981)]). [Proc. Natl. Acad. Sci. U.S.A. 78, 2169-2173 (1981)] also sequenced S.typhimurium trpA region. [Nucleic Acids Res. 9, 6647-6668 (1981)] compiles sequences from [J. Mol. Biol. 121, 113-137 (1978)],[Nature 277, 486-489 (1979)], [Proc. Natl. Acad. Sci. U.S.A. 76, 5244-5248 (1979)],[J. Mol. Biol. 142, 519-530 (1980)],[J. Mol. Biol. 142, 489-502 (1980)],[J. Mol. Biol. 142, 503-517 (1980)],[J. Mol. Biol. 146, 45-54 (1981)],[J. Mol. Biol. 156, 245-256 (1982)]. FEATURES Location/Qualifiers -35_signal 285..290 protein_bind 301..318 /bound_moiety="trpR regulatory protein" /evidence=EXPERIMENTAL -10_signal 309..315 mRNA 321..461 /note="trp mRNA (alt.) [Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976)],[J. Mol. Biol. 103, 351-381 (1976)],[J. Mol. Biol. 121, 113-137 (1978)],[Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979)],[Nucleic Acids Res. 10, 6639-6657 (1982)]" mRNA 321..7046 /note="trp mRNA (alt.) [Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976)],[J. Mol. Biol. 103, 351-381 (1976)], [Proc. Natl. Acad. Sci. U.S.A. 75, 5442-5446 (1978)],[J. Mol. Biol. 121, 113-137 (1978)],[Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (" CDS 347..391 /note="trp operon leader peptide (putative)" /codon_start=347 CDS 483..2045 /note="anthranilate synthetase component I /nomgen='trpE'" /codon_start=483 CDS 2045..3640 /note="anthranilate synthetase component II: glutamine amidotransferase and phosphoribosyl anthranilate synthetase /nomgen='trpD'" /codon_start=2045 CDS 3644..5002 /note="anthranilate isomerase /nomgen='trpC'" /codon_start=3644 CDS 5413..6207 /note="tryptophan synthetase beta subunit /nomgen='trpB'" /codon_start=5413 CDS 6207..7013 /note="tryptophan synthetase alpha subunit /nomgen='trpA'" /codon_start=6207 BASE COUNT 1779 a 1980 c 2022 g 1754 t 4 others ORIGIN 9 bp upstream from HhaI site [J. Mol. Biol. 121, 113-137 (1978)]. 1 ccgggaataa gattcaacgc cagtcccgaa cgtgaaattt cctctcttgc tggcgcgatt 61 gcagctgtgg tgtcatggtc ggtgatcgcc agggtgccga cgcgcatctc gactgcacgg 121 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg 181 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg 241 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc 301 gaactagtta actagtacgc aagttcacgt aaaaagggta tcgacaatga aagcaatttt 361 cgtactgaaa ggttggtggc gcacttcctg aaacgggcag tgtattcacc atgcgtaaag 421 caatcagata cccagcccgc ctaatgagcg ggcttttttt tgaacaaaat tagagaataa 481 caatgcaaac acaaaaaccg actctcgaac tgctaacctg cgaaggcgct tatcgcgaca 541 atcccaccgc gctttttcac cagttgtgtg gggatcgtcc ggcaacgctg ctgctggaat 601 ccgcagatat cgacagcaaa gatgatttaa aaagcctgct gctggtagac agtgcgctgc 661 gcattacagc tttaggtgac actgtcacaa tccaggcact ttccggcaac ggcgaagccc 721 tcctggcact actggataac gccctgcctg cgggtgtgga aagtgaacaa tcaccaaact 781 gccgtgtgct gcgcttcccc cctgtcagtc cactgctgga tgaagacgcc cgcttatgct 841 ccctttcggt ttttgacgct ttccgtttat tgcagaatct gttgaatgta ccgaaggaag 901 aacgagaagc catgttcttc agcggcctgt tctcttatga ccttgtggcg ggatttgaag 961 atttaccgca actgtcagcg gaaaataact gccctgattt ctgtttttat ctcgctgaaa 1021 cgctgatggt gattgaccat cagaaaaaaa gcacccgtat tcaggccagc ctgtttgctc 1081 cgaatgaaga agaaaaacaa cgtctcactg ctcgcctgaa cgaactacgt cagcaactga 1141 ccgaagccgc gccgccgctg ccagtggttt ccgtgccgca tatgcgttgt gaatgtaatc 1201 agagcgatga agagttcggt ggcgtagtgc gtttgttgca aaaagcgatt cgcgctggag 1261 aaattttcca ggtggtgcca tctcgccgtt tctctctgcc ctgcccgtca ccgctggcgg 1321 cctattacgt gctgaaaaag agtaatccca gcccgtacat gttttttatg caggataatg 1381 atttcaccct atttggcgcg tcgccggaaa gctcgctcaa gtatgatgcc accagccgcc 1441 agattgagat ctacccgatt gccggaacac gcccacgcgg tcgtcgcgcc gatggttcac 1501 tggacagaga tctcgacagc cgtattgaac tggaaatgcg taccgatcat aaagagctgt 1561 ctgaacatct gatgctggtt gatctcgccc gtaatgatct ggcacgcatt tgcacccccg 1621 gcagccgcta cgtcgccgat ctcaccaaag ttgaccgtta ttcctatgtg atgcacctcg 1681 tctctcgcgt agtcggcgaa ctgcgtcacg atcttgacgc cctgcacgct tatcgcgcct 1741 gtatgaatat ggggacgtta agcggtgcgc cgaaagtacg cgctatgcag ttaattgccg 1801 aggcggaagg tcgtcgccgc ggcagctacg gcggcgcggt aggttatttc accgcgcatg 1861 gcgatctcga cacctgcatt gtgatccgct cggcgctggt ggaaaacggt atcgccaccg 1921 tgcaagcggg tgctggtgta gtccttgatt ctgttccgca gtcggaagcc gacgaaaccc 1981 gtaacaaagc ccgcgctgta ctgcgcgcta ttgccaccgc gcatcatgca caggagactt 2041 tctgatggct gacattctgc tgctcgataa tatcgactct tttacgtaca acctggcaga 2101 tcagttgcgc agcaatgggc ataacgtggt gatttaccgc aaccatatac cggcgcaaac 2161 cttaattgaa cgcttggcga ccatgagtaa tccggtgctg atgctttctc ctggccccgg 2221 tgtgccgagc gaagccggtt gtatgccgga actcctcacc cgcttgcgtg gcaagctgcc 2281 cattattggc atttgcctcg gacatcaggc gattgtcgaa gcttacgggg gctatgtcgg 2341 tcaggcgggc gaaattctcc acggtaaagc ctccagcatt gaacatgacg gtcaggcgat 2401 gtttgccgga ttaacaaacc cgctgccggt ggcgcgttat cactcgctgg ttggcagtaa 2461 cattccggcc ggtttaacca tcaacgccca ttttaatggc atggtgatgg cagtacgtca 2521 cgatgcggat cgcgtttgtg gattccagtt ccatccggaa tccattctca ccacccaggg 2581 cgctcgcctg ctggaacaaa cgctggcctg ggcgcagcat aaactagagc cagccaacac 2641 gctgcaaccg attctggaaa aactgtatca ggcgcagacg cttagccaac aagaaagcca 2701 ccagctgttt tcagcggtgg tgcgtggcga gctgaagccg gaacaactgg cggcggcgct 2761 ggtgagcatg aaaattcgcg gtgagcaccc gaacgagatc gccggggcag caaccgcgct 2821 actggaaaac gcagcgccgt tcccgcgccc ggattatctg tttgctgata tcgtcggtac 2881 tggcggtgac ggcagcaaca gtatcaatat ttctaccgcc agtgcgtttg tcgccgcggc 2941 ctgtgggctg aaagtggcga aacacggcaa ccgtagcgtc tccagtaaat ctggttcgtc 3001 cgatctgctg gcggcgttcg gtattaatct tgatatgaac gccgataaat cgcgccaggc 3061 gctggatgag ttaggtgtat gtttcctctt tgcgccgaag tatcacaccg gattccgcca 3121 cgcgatgccg gttcgccagc aactgaaaac ccgcaccctg ttcaatgtgc tggggccatt 3181 gattaacccg gcgcatccgc cgctggcgtt aattggtgtt tatagtccgg aactggtgct 3241 gccgattgcc gaaaccttgc gcgtgctggg gtatcaacgc gcggcggtgg tgcacagcgg 3301 cgggatggat gaagtttcat tacacgcgcc gacaatcgtt gccgaactgc atgacggcga 3361 aattaaaagc tatcagctca ccgcagaaga ctttggcctg acaccctacc accaggagca 3421 actggcaggc ggaacaccgg aagaaaaccg tgacatttta acacgtttgt tacaaggtaa 3481 aggcgacgcc gcccatgaag cagccgtcgc tgcgaacgtc gccatgttaa tgcgcctgca 3541 tggccatgaa gatctgcaag ccaatgcgca aaccgttctt gaggtactgc gcagtggttc 3601 cgcttacgac agagtcaccg cactggcggc acgagggtaa atgatgcaaa ccgttttagc 3661 gaaaatcgtc gcagacaagg cgatttgggt agaagcccgc aaacagcagc aaccgctggc 3721 cagttttcag aatgaggttc agccgagcac gcgacatttt tatgatgcgc tacagggtgc 3781 gcgcacggcg tttattctgg agtgcaagaa agcgtcgccg tcaaaaggcg tgatccgtga 3841 tgatttcgat ccagcacgca ttgccgccat ttataaacat tacgcttcgg caatttcggt 3901 gctgactgat gagaaatatt tcaggggtag ctttaatttc ctccccatcg tcagccaaat 3961 cgccccgcag ccgattttat gtaaagactt cattatcgac ccttaccaga tctatctggc 4021 gcgctattac caggccgatg cctgcttatt aatgctttca gtactggatg acgaccaata 4081 tcgccagctt gccgccgtcg ctcacagtct ggagatgggg gtgctgaccg aagtcagtaa 4141 tgaagaggaa caggagcgcg ccattgcatt gggagcaaag gtcgttggca tcaacaaccg 4201 cgatctgcgt gatttgtcga ttgatctcaa ccgtacccgc gagcttgcgc cgaaactggg 4261 gcacaacgtg acggtaatca gcgaatccgg catcaatact tacgctcagg tgcgcgagtt 4321 aagccacttc gctaacggtt ttctgattgg ttcggcgttg atggcccatg acgatttgca 4381 cgccgccgtg cgccgggtgt tgctgggtga gaataaagta tgtggcctga cgcgtgggca 4441 agatgctaaa gcagcttatg acgcgggcgc gatttacggt gggttgattt ttgttgcgac 4501 atcaccgcgt tgcgtcaacg ttgaacaggc gcaggaagtg atggctgcgg caccgttgca 4561 gtatgttggc gtgttccgca atcacgatat tgccgatgtg gtggacaaag ctaaggtgtt 4621 atcgctggtg gcagtgcaac tgcatggtaa tgaagaacag ctgtatatcg atacgctgcg 4681 tgaagctctg ccagcacatg ttgccatctg gaaagcatta agcgtcggtg aaaccctgcc 4741 cgcccgcgag tttcagcacg ttgataaata tgttttagac aacggccagg gtggaagcgg 4801 gcaacgtttt gactggtcac tattaaatgg tcaaacgctt ggcaacgttc tgctggcggg 4861 gggcttaggc gcagataact gcgtggaagc ggcacaaacc ggctgcgccg gacttgattt 4921 taattctgct gtagagtcgc aaccgggcat caaagacgca cgtcttttgg cctcggtttt 4981 ccagacgctg cgcgcatatt aaggaaagga acaatgacaa cattacttaa cccctatttt 5041 ggtgagtttg gcggcatgta cgtgccacaa atcctgatgc ctgctctgcg ccagctggaa 5101 gaagcttttg tcagtgcgca aaaagatcct gaatttcagg ctcagttcaa cgacctgctg 5161 aaaaactatg ccgggcgtcc aaccgcgctg accaaatgcc agaacattac agccgggacg 5221 aacaccacgc tgtatctcaa gcgtgaagat ttgctgcacg gcggcgcgca taaaactaac 5281 caggtgctgg ggcaggcgtt gctggcgaag cggatgggta aaaccgaaat catcgccgaa 5341 accggtgccg gtcagcatgg cgtggcgtcg gccctggcca gcgccctgct cggcctgaaa 5401 tgccgtattt atatgggtgc caaagacgtt gaacgccagt cgcctaacgt ttttcgtatg 5461 cgcttaatgg gtgcggaagt gatcccggtg catagcggtt ccgcgacgct gaaagatgcc 5521 tgtaacgagg cgctgcgcga ctggtccggt agttacgaaa ccgcgcacta tatgctgggc 5581 accgcagctg gcccgcatcc ttatccgacc attgtgcgtg agtttcagcg gatgattggc 5641 gaagaaacca aagcgcagat tctggaaaga gaaggtcgcc tgccggatgc cgttatcgcc 5701 tgtgttggcg gcggttcgaa tgccatcggc atgtttgctg atttcatcaa tgaaaccaac 5761 gtcggcctga ttggtgtgga gccaggtggt cacggtatcg aaactggcga gcacggcgca 5821 ccgctaaaac atggtcgcgt gggtatctat ttcggtatga aagcgccgat gatgcaaacc 5881 gaagacgggc agattgaaga atcttactcc atctccgccg gactggattt cccgtctgtc 5941 ggcccacaac acgcgtatct taacagcact ggacgcgctg attacgtgtc tattaccgat 6001 gatgaagccc ttgaagcctt caaaacgctg tgcctgcacg aagggatcat cccggcgctg 6061 gaatcctccc acgccttggc ccatgcgttg aaaatgatgc gcgaaaaccc ggataaagag 6121 cagctactgg tggttaacct ttccggtcgc ggcgataaag acatcttcac cgttcacgat 6181 attttgaaag cacgagggga aatctgatgg aacgctacga atctctgttt gcccagttga 6241 aggagcgcaa agaaggcgca ttcgttcctt tcgtcacgct cggtgatccg ggcattgagc 6301 agtcattgaa aattatcgat acgctaattg aagccggtgc tgacgcgctg gagttaggta 6361 tccccttctc cgacccactg gcggatggcc cgacgattca aaacgccact ctgcgcgcct 6421 ttgcggcagg tgtgactccg gcacaatgtt ttgaaatgct ggcactgatt cgccagaaac 6481 acccgaccat tcccattggc ctgttgatgt atgccaatct ggtgtttaac aaaggcattg 6541 atgagtttta tgcccagtgc gaaaaagtcg gcgtcgattc ggtgctggtt gccgatgtgc 6601 cagttgaaga gtccgcgccc ttccgccagg ccgcgttgcg tcacaacgtc gcacctatct 6661 tcatctgccc gccaaatgcc gatgacgacc tgctgcgcca gatagcctct tacggtcgtg 6721 gttacaccta tttgctgtca cgagcaggcg tgaccggcgc agaaaaccgc gccgcgttac 6781 ccctcaatca tctggttgcg aagctgaaag agtacaacgc tgcacctcca ttgcagggat 6841 ttggtatttc cgccccggat caggtaaaag cagcgattga tgcaggagct gcgggcgcga 6901 tttctggttc ggccattgtt aaaatcatcg agcaacatat taatgagcca gagaaaatgc 6961 tggcggcact gaaagttttt gtacaaccga tgaaagcggc gacgcgcagt taatcccaca 7021 gccgccagtt ccgctggcgg cattttaact ttctttaatg aagccggaaa aatcctaaat 7081 tcatttaata tttatctttt taccgtttcg cttaccccgg tcgatcgtyr acttacgtca 7141 tttttccgcc caacagtaat ataaacaaac aaattaaacc cgcaacataa caccagtaaa 7201 atcaataatt ttctctaagt cacttattcc tcaggtaatt cttaatatat ccagaatgtt 7261 cctcaaaata tattttccct ctatcttctc gttgcgctta atttgactaa ttctcattag 7321 cgactaattt taatgagtgt cgacacacaa cactcatatt aatgaaacaa tgcaacgcaa 7381 cgggagaaat aacatggccg aacatcgtgg tggttcagga aatttcgccg aagaccgtga 7441 gaaggcatcc gacgcagccg taaaggcggt cagcatagcg gcggtaattt taaaaatgat 7501 cgcaacgcgc atctgaagcg ggtaaaaaag gcggtyrac //
GenBank-Updates@genbank.bio.net (05/14/91)
LOCUS ECOTGP 7539 bp ds-DNA BCT 14-MAY-1991 DEFINITION E.coli tryptophan operon: entire DNA sequence. ACCESSION J01714 M12471 M12472 M25593 M59208 KEYWORDS anthranilate isomerase; anthranilate synthetase; attenuator; glutamine amidotransferase; isomerase; leader peptide; phosphoribosyl anthranilate synthetase; synthetase; transferase; trp operon; trpA gene; trpB gene; trpC gene; trpD gene; trpE gene; tryptophan synthetase. SOURCE Escherichia coli RNA and DNA. ORGANISM Escherichia coli Prokaryota; Bacteria; Gracilicutes; Scotobacteria; Facultatively anaerobic rods; Enterobacteriaceae. REFERENCE 1 (bases 5917 to 6133) AUTHORS Platt,T. and Yanofsky,C. TITLE An intercistronic region and ribosome-binding site in bacterial messenger RNA JOURNAL Proc. Natl. Acad. Sci. U.S.A. 72, 2399-2403 (1975) STANDARD full staff_review REFERENCE 2 (bases 84 to 141) AUTHORS Bennett,G.N., Schweingruber,M.E., Brown,K.D., Squires,C. and Yanofsky,C. TITLE Nucleotide sequence of region preceding trp mRNA initiation site and its role in promoter and operator function JOURNAL Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976) STANDARD full staff_review REFERENCE 3 (bases 117 to 310) AUTHORS Squires,C., Lee,F., Bertrand,K., Squires,C.L., Bronson,M.J. and Yanofsky,C. TITLE Nucleotide sequence of the 5' end of tryptophan messenger RNA of Escherichia coli JOURNAL J. Mol. Biol. 103, 351-381 (1976) STANDARD full staff_review REFERENCE 4 (bases 230 to 272) AUTHORS Bertrand,K., Korn,L.J., Lee,F. and Yanofsky,C. TITLE The attenuator of the tryptophan operon of Escherichia coli: heterogeneous 3'-OH termini in vivo and deletion mapping of functions JOURNAL J. Mol. Biol. 117, 227-247 (1977) STANDARD full staff_review REFERENCE 5 (bases 230 to 272) AUTHORS Stauffer,G.V., Zurawski,G. and Yanofsky,C. TITLE Single base-pair alterations in the Escherichia coli trp operon leader region that relieve transcription termination at the trp attenuator JOURNAL Proc. Natl. Acad. Sci. U.S.A. 75, 4833-4837 (1978) STANDARD full staff_review REFERENCE 6 (bases 6707 to 6863) AUTHORS Wu,A.M. and Platt,T. TITLE Transcription termination: nucleotide sequence at 3' end of tryptophan operon in Escherichia coli JOURNAL Proc. Natl. Acad. Sci. U.S.A. 75, 5442-5446 (1978) STANDARD full staff_review REFERENCE 7 (bases 0 to 0) AUTHORS Bennett,G.N., Schweingruber,M.E., Brown,K.D., Squires,C. and Yanofsky,C. TITLE Nucleotide sequence of the promoter-operator region of the tryptophan operon of Escherichia coli JOURNAL J. Mol. Biol. 121, 113-137 (1978) STANDARD full staff_review REFERENCE 8 (bases 36 to 136) AUTHORS Brown,K.D., Bennet,G.N., Lee,F., Schweingruber,M.E. and Yanofsky,C. TITLE RNA polymerase interaction at the promoter-operator region of the tryptophan operon of Escherichia coli and Salmonella typhimurium JOURNAL J. Mol. Biol. 121, 153-177 (1978) STANDARD simple staff_entry REFERENCE 9 (bases 2351 to 2503) AUTHORS Miozzari,G.F. and Yanofsky,C. TITLE Gene fusion during the evolution of the tryptophan operon in enterobacteriaceae JOURNAL Nature 277, 486-489 (1979) STANDARD full staff_review REFERENCE 10 (bases 5932 to 6809) AUTHORS Nichols,B.P. and Yanofsky,C. TITLE Nucleotide sequences of trpA of Salmonella typhimurium JOURNAL Proc. Natl. Acad. Sci. U.S.A. 76, 5244-5248 (1979) STANDARD full staff_review REFERENCE 11 (bases 117 to 256) AUTHORS Oxender,D.L., Zurawski,G. and Yanofsky,C. TITLE Attenuation in the Escherichia coli tryptophan operon: role of RNA secondary structure involving the tryptophan codon region JOURNAL Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979) STANDARD full staff_review REFERENCE 12 (bases 6707 to 7335) AUTHORS Wu,A.M., Chapman,A.B., Platt,T., Guarente,L.P. and Beckwith,J. TITLE Deletions of distal sequence affect termination of transcription at the end of the tryptophan operon in E. coli JOURNAL Cell 19, 829-836 (1980) STANDARD full staff_review REFERENCE 13 (bases 230 to 296) AUTHORS Farnham,P.J. and Platt,T. TITLE A model for transcription termination suggested by studies on the trp attenuator in vitro using base analogs JOURNAL Cell 20, 739-748 (1980) STANDARD full staff_review REFERENCE 14 (bases 4810 to 6003) AUTHORS Crawford,I.P., Nichols,B.P. and Yanofsky,C. TITLE Nucleotide sequence of the trpB gene in Escherichia coli and Salmonella typhimurium JOURNAL J. Mol. Biol. 142, 489-502 (1980) STANDARD full staff_review REFERENCE 15 (bases 1761 to 2443) AUTHORS Nichols,B.P., Miozzari,G.F., van Cleemput,M., Bennett,G.N. and Yanofsky,C. TITLE Nucleotide sequences of the trpG regions of Escherichia coli, Shigella dysenteriae, Salmonella typhimurium and Serratia marcescens JOURNAL J. Mol. Biol. 142, 503-517 (1980) STANDARD full staff_review REFERENCE 16 (bases 3422 to 4824) AUTHORS Christie,G.E. and Platt,T. TITLE Gene structure in the tryptophan operon of Escherichia coli: nucleotide sequence of trpC and the flanking intercistronic regions JOURNAL J. Mol. Biol. 142, 519-530 (1980) STANDARD full staff_review REFERENCE 17 (bases 5932 to 6809) AUTHORS Schneider,W.P., Nichols,B.P. and Yanofsky,C. TITLE Procedure for production of hybrid genes and proteins and its use in assessing significance of amino acid differences in homologous tryptophan synthetase alpha polypeptides JOURNAL Proc. Natl. Acad. Sci. U.S.A. 78, 2169-2173 (1981) STANDARD full staff_review REFERENCE 18 (bases 6807 to 6856; 7057 to 7119) AUTHORS Wu,A.M., Christie,G.E. and Platt,T. TITLE Tandem termination sites in the tryptophan operon of Escherichia coli JOURNAL Proc. Natl. Acad. Sci. U.S.A. 78, 2913-2917 (1981) STANDARD full staff_review REFERENCE 19 (bases 279 to 1843) AUTHORS Nichols,B.P., van Cleemput,M. and Yanofsky,C. TITLE Nucleotide sequence of Escherichia coli trpE: anthranilate synthetase component I contains no tryptophan residues JOURNAL J. Mol. Biol. 146, 45-54 (1981) STANDARD full staff_review REFERENCE 20 (sites) AUTHORS Yanofsky,C., Platt,T., Crawford,I.P., Nichols,B.P., Christie,G.E., Horowitz,H., van Cleemput,M. and Wu,A.M. TITLE The complete nucleotide sequence of the tryptophan operon of Escherichia coli JOURNAL Nucleic Acids Res. 9, 6647-6668 (1981) STANDARD full staff_review REFERENCE 21 (bases 2504 to 3436) AUTHORS Horowitz,H., Christie,G.E. and Platt,T. TITLE Nucleotide sequence of the trpD gene, encoding anthranilate synthetase component II of Escherichia coli JOURNAL J. Mol. Biol. 156, 245-256 (1982) STANDARD full staff_review REFERENCE 22 (bases 57 to 137) AUTHORS Windass,J.D., Newton,C.R., De Maeyer-Guignard,J., Moore,V.E., Markham,A.F. and Edge,M.D. TITLE The construction of a synthetic Escherichia coli trp promoter and its use in the expression of a synthetic interferon gene JOURNAL Nucleic Acids Res. 10, 6639-6657 (1982) STANDARD full staff_review REFERENCE 23 (sites) AUTHORS Kolter,R. and Yanofsky,C. TITLE Genetic analysis of the tryptophan operon regulatory region using site-directed mutagenesis JOURNAL J. Mol. Biol. 175, 299-312 (1984) STANDARD full staff_review REFERENCE 24 (bases 1 to 350) AUTHORS Kane,J.F., Balaban,S.M. and Bogosian,G. TITLE Commercial production of bovine somatotropin in Escherichia coli JOURNAL (in) Sikes,C.S. and Wheeler,A.P. (Eds.); Surface reactive peptides and polymers. Discovery and commercialization.: In press, American Chemical Society, Washington, D.C. (1990) STANDARD simple staff_entry COMMENT [Nucleic Acids Res. 9, 6647-6668 (1981)] review; bases 77 to 6809; compiled. [J. Mol. Biol. 175, 299-312 (1984)] sites; mutational analysis of the regulatory region. The tryptophan operon of E.coli consists of a repressor(trpR), a promoter(trpP), an operator(trpO), an attenuator which is part of a leader peptide region(trpL) and five structural genes: trpE(anthranilate synthetase), trpD(glutamine amido transferase and anthranilate 5-phosphoribosylpyrophosphate phosphoribosyl- transferase), trpC(phosphoribosyl anthranilate isomerase-indole glycerol phosphate synthetase), trpB(tryptophan synthetase beta) and trpA(tryptophan synthetase alpha). The promoter region covers approximately 40 bases upstream from the mRNA initiation site(75-116); the operator approximately 20 bases upstream with two-fold axes of symmetry around 104-105 and 109-110([Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976)],[J. Mol. Biol. 121, 113-137 (1978)],[J. Mol. Biol. 156, 245-256 (1982)]). The attenuator region is the first 140 nucleotides(117-256) of the mRNA leader, a G-C rich region with a two-fold axis of symmetry around base 240 and an A-T rich region with its axis about bases 259-260; it provides a second site for control of transcription ([J. Mol. Biol. 117, 227-247 (1977)], [Proc. Natl. Acad. Sci. U.S.A. 75, 4833-4837 (1978)],[Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979)],[Cell 20, 739-748 (1980)]). Two mRNA termination regions are reported: trpT (bases 6807-6856) and trpT' (bases 7057-7119), the first of which bears some similarity to the attenuator region ([Proc. Natl. Acad. Sci. U.S.A. 78, 2913-2917 (1981)]). A chi site for recombination is localized between bases 2492 and 2501 and the trp-P2 promoter is located between bases 3240 and 3280 ([J. Mol. Biol. 156, 245-256 (1982)]). The trpE gene is unusual in that it codes for no tryptophan residues([J. Mol. Biol. 146, 45-54 (1981)]). The two enzymatic functions coded by trpG and trpD genes in S.marcescens are coded by the single trpD gene in E.coli and other enterobacteriaceae. This appears to have occurred via base changes at sites 2420 and 2438. The intercistronic regions for the structural genes show little superfluity: the trpE-trpD and trpB-trpA boundaries consist of 'tgatg'; the trpD-trpC boundary is 'taaatgatg' and the trpC-trpB boundary is 'taaggaaaggaacaatg'. All the cistrons show a high degree of homology with their correlates among the enterobacteriaceae. Sequence discrepancies in early work([J. Mol. Biol. 103, 351-381 (1976)]) are corrected in later work from the same laboratory([Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979)], [Nucleic Acids Res. 9, 6647-6668 (1981)]). [Proc. Natl. Acad. Sci. U.S.A. 78, 2169-2173 (1981)] also sequenced S.typhimurium trpA region. [Nucleic Acids Res. 9, 6647-6668 (1981)] compiles sequences from [J. Mol. Biol. 121, 113-137 (1978)],[Nature 277, 486-489 (1979)], [Proc. Natl. Acad. Sci. U.S.A. 76, 5244-5248 (1979)],[J. Mol. Biol. 142, 519-530 (1980)],[J. Mol. Biol. 142, 489-502 (1980)],[J. Mol. Biol. 142, 503-517 (1980)],[J. Mol. Biol. 146, 45-54 (1981)],[J. Mol. Biol. 156, 245-256 (1982)]. FEATURES Location/Qualifiers -35_signal 285..290 protein_bind 301..318 /bound_moiety="trpR regulatory protein" /evidence=EXPERIMENTAL -10_signal 309..315 mRNA 321..461 /note="trp mRNA (alt.) [Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976)],[J. Mol. Biol. 103, 351-381 (1976)],[J. Mol. Biol. 121, 113-137 (1978)],[Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (1979)],[Nucleic Acids Res. 10, 6639-6657 (1982)]" mRNA 321..7046 /note="trp mRNA (alt.) [Proc. Natl. Acad. Sci. U.S.A. 73, 2351-2355 (1976)],[J. Mol. Biol. 103, 351-381 (1976)], [Proc. Natl. Acad. Sci. U.S.A. 75, 5442-5446 (1978)],[J. Mol. Biol. 121, 113-137 (1978)],[Proc. Natl. Acad. Sci. U.S.A. 76, 5524-5528 (" CDS 347..391 /note="trp operon leader peptide (putative)" /codon_start=347 CDS 483..2045 /note="anthranilate synthetase component I /nomgen='trpE'" /codon_start=483 old_sequence 1991..1991 /location=J. Mol. Biol. 142, 503-517 (1980) | 27..27 old_sequence 1997..1997 /location=J. Mol. Biol. 142, 503-517 (1980) | 33..33 CDS 2045..3640 /note="anthranilate synthetase component II: glutamine amidotransferase and phosphoribosyl anthranilate synthetase /nomgen='trpD'" /codon_start=2045 CDS 3644..5002 /note="anthranilate isomerase /nomgen='trpC'" /codon_start=3644 CDS 5014..6207 /note="tryptophan synthetase beta subunit /nomgen='trpB'" /codon_start=5014 conflict 6153..6153 /location=Proc. Natl. Acad. Sci. U.S.A. 78, 2169-2173 (1981) | 18..18 CDS 6207..7013 /note="tryptophan synthetase alpha subunit /nomgen='trpA'" /codon_start=6207 BASE COUNT 1779 a 1980 c 2022 g 1754 t 4 others ORIGIN 9 bp upstream from HhaI site [J. Mol. Biol. 121, 113-137 (1978)]. 1 ccgggaataa gattcaacgc cagtcccgaa cgtgaaattt cctctcttgc tggcgcgatt 61 gcagctgtgg tgtcatggtc ggtgatcgcc agggtgccga cgcgcatctc gactgcacgg 121 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg 181 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg 241 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc 301 gaactagtta actagtacgc aagttcacgt aaaaagggta tcgacaatga aagcaatttt 361 cgtactgaaa ggttggtggc gcacttcctg aaacgggcag tgtattcacc atgcgtaaag 421 caatcagata cccagcccgc ctaatgagcg ggcttttttt tgaacaaaat tagagaataa 481 caatgcaaac acaaaaaccg actctcgaac tgctaacctg cgaaggcgct tatcgcgaca 541 atcccaccgc gctttttcac cagttgtgtg gggatcgtcc ggcaacgctg ctgctggaat 601 ccgcagatat cgacagcaaa gatgatttaa aaagcctgct gctggtagac agtgcgctgc 661 gcattacagc tttaggtgac actgtcacaa tccaggcact ttccggcaac ggcgaagccc 721 tcctggcact actggataac gccctgcctg cgggtgtgga aagtgaacaa tcaccaaact 781 gccgtgtgct gcgcttcccc cctgtcagtc cactgctgga tgaagacgcc cgcttatgct 841 ccctttcggt ttttgacgct ttccgtttat tgcagaatct gttgaatgta ccgaaggaag 901 aacgagaagc catgttcttc agcggcctgt tctcttatga ccttgtggcg ggatttgaag 961 atttaccgca actgtcagcg gaaaataact gccctgattt ctgtttttat ctcgctgaaa 1021 cgctgatggt gattgaccat cagaaaaaaa gcacccgtat tcaggccagc ctgtttgctc 1081 cgaatgaaga agaaaaacaa cgtctcactg ctcgcctgaa cgaactacgt cagcaactga 1141 ccgaagccgc gccgccgctg ccagtggttt ccgtgccgca tatgcgttgt gaatgtaatc 1201 agagcgatga agagttcggt ggcgtagtgc gtttgttgca aaaagcgatt cgcgctggag 1261 aaattttcca ggtggtgcca tctcgccgtt tctctctgcc ctgcccgtca ccgctggcgg 1321 cctattacgt gctgaaaaag agtaatccca gcccgtacat gttttttatg caggataatg 1381 atttcaccct atttggcgcg tcgccggaaa gctcgctcaa gtatgatgcc accagccgcc 1441 agattgagat ctacccgatt gccggaacac gcccacgcgg tcgtcgcgcc gatggttcac 1501 tggacagaga tctcgacagc cgtattgaac tggaaatgcg taccgatcat aaagagctgt 1561 ctgaacatct gatgctggtt gatctcgccc gtaatgatct ggcacgcatt tgcacccccg 1621 gcagccgcta cgtcgccgat ctcaccaaag ttgaccgtta ttcctatgtg atgcacctcg 1681 tctctcgcgt agtcggcgaa ctgcgtcacg atcttgacgc cctgcacgct tatcgcgcct 1741 gtatgaatat ggggacgtta agcggtgcgc cgaaagtacg cgctatgcag ttaattgccg 1801 aggcggaagg tcgtcgccgc ggcagctacg gcggcgcggt aggttatttc accgcgcatg 1861 gcgatctcga cacctgcatt gtgatccgct cggcgctggt ggaaaacggt atcgccaccg 1921 tgcaagcggg tgctggtgta gtccttgatt ctgttccgca gtcggaagcc gacgaaaccc 1981 gtaacaaagc ccgcgctgta ctgcgcgcta ttgccaccgc gcatcatgca caggagactt 2041 tctgatggct gacattctgc tgctcgataa tatcgactct tttacgtaca acctggcaga 2101 tcagttgcgc agcaatgggc ataacgtggt gatttaccgc aaccatatac cggcgcaaac 2161 cttaattgaa cgcttggcga ccatgagtaa tccggtgctg atgctttctc ctggccccgg 2221 tgtgccgagc gaagccggtt gtatgccgga actcctcacc cgcttgcgtg gcaagctgcc 2281 cattattggc atttgcctcg gacatcaggc gattgtcgaa gcttacgggg gctatgtcgg 2341 tcaggcgggc gaaattctcc acggtaaagc ctccagcatt gaacatgacg gtcaggcgat 2401 gtttgccgga ttaacaaacc cgctgccggt ggcgcgttat cactcgctgg ttggcagtaa 2461 cattccggcc ggtttaacca tcaacgccca ttttaatggc atggtgatgg cagtacgtca 2521 cgatgcggat cgcgtttgtg gattccagtt ccatccggaa tccattctca ccacccaggg 2581 cgctcgcctg ctggaacaaa cgctggcctg ggcgcagcat aaactagagc cagccaacac 2641 gctgcaaccg attctggaaa aactgtatca ggcgcagacg cttagccaac aagaaagcca 2701 ccagctgttt tcagcggtgg tgcgtggcga gctgaagccg gaacaactgg cggcggcgct 2761 ggtgagcatg aaaattcgcg gtgagcaccc gaacgagatc gccggggcag caaccgcgct 2821 actggaaaac gcagcgccgt tcccgcgccc ggattatctg tttgctgata tcgtcggtac 2881 tggcggtgac ggcagcaaca gtatcaatat ttctaccgcc agtgcgtttg tcgccgcggc 2941 ctgtgggctg aaagtggcga aacacggcaa ccgtagcgtc tccagtaaat ctggttcgtc 3001 cgatctgctg gcggcgttcg gtattaatct tgatatgaac gccgataaat cgcgccaggc 3061 gctggatgag ttaggtgtat gtttcctctt tgcgccgaag tatcacaccg gattccgcca 3121 cgcgatgccg gttcgccagc aactgaaaac ccgcaccctg ttcaatgtgc tggggccatt 3181 gattaacccg gcgcatccgc cgctggcgtt aattggtgtt tatagtccgg aactggtgct 3241 gccgattgcc gaaaccttgc gcgtgctggg gtatcaacgc gcggcggtgg tgcacagcgg 3301 cgggatggat gaagtttcat tacacgcgcc gacaatcgtt gccgaactgc atgacggcga 3361 aattaaaagc tatcagctca ccgcagaaga ctttggcctg acaccctacc accaggagca 3421 actggcaggc ggaacaccgg aagaaaaccg tgacatttta acacgtttgt tacaaggtaa 3481 aggcgacgcc gcccatgaag cagccgtcgc tgcgaacgtc gccatgttaa tgcgcctgca 3541 tggccatgaa gatctgcaag ccaatgcgca aaccgttctt gaggtactgc gcagtggttc 3601 cgcttacgac agagtcaccg cactggcggc acgagggtaa atgatgcaaa ccgttttagc 3661 gaaaatcgtc gcagacaagg cgatttgggt agaagcccgc aaacagcagc aaccgctggc 3721 cagttttcag aatgaggttc agccgagcac gcgacatttt tatgatgcgc tacagggtgc 3781 gcgcacggcg tttattctgg agtgcaagaa agcgtcgccg tcaaaaggcg tgatccgtga 3841 tgatttcgat ccagcacgca ttgccgccat ttataaacat tacgcttcgg caatttcggt 3901 gctgactgat gagaaatatt tcaggggtag ctttaatttc ctccccatcg tcagccaaat 3961 cgccccgcag ccgattttat gtaaagactt cattatcgac ccttaccaga tctatctggc 4021 gcgctattac caggccgatg cctgcttatt aatgctttca gtactggatg acgaccaata 4081 tcgccagctt gccgccgtcg ctcacagtct ggagatgggg gtgctgaccg aagtcagtaa 4141 tgaagaggaa caggagcgcg ccattgcatt gggagcaaag gtcgttggca tcaacaaccg 4201 cgatctgcgt gatttgtcga ttgatctcaa ccgtacccgc gagcttgcgc cgaaactggg 4261 gcacaacgtg acggtaatca gcgaatccgg catcaatact tacgctcagg tgcgcgagtt 4321 aagccacttc gctaacggtt ttctgattgg ttcggcgttg atggcccatg acgatttgca 4381 cgccgccgtg cgccgggtgt tgctgggtga gaataaagta tgtggcctga cgcgtgggca 4441 agatgctaaa gcagcttatg acgcgggcgc gatttacggt gggttgattt ttgttgcgac 4501 atcaccgcgt tgcgtcaacg ttgaacaggc gcaggaagtg atggctgcgg caccgttgca 4561 gtatgttggc gtgttccgca atcacgatat tgccgatgtg gtggacaaag ctaaggtgtt 4621 atcgctggtg gcagtgcaac tgcatggtaa tgaagaacag ctgtatatcg atacgctgcg 4681 tgaagctctg ccagcacatg ttgccatctg gaaagcatta agcgtcggtg aaaccctgcc 4741 cgcccgcgag tttcagcacg ttgataaata tgttttagac aacggccagg gtggaagcgg 4801 gcaacgtttt gactggtcac tattaaatgg tcaaacgctt ggcaacgttc tgctggcggg 4861 gggcttaggc gcagataact gcgtggaagc ggcacaaacc ggctgcgccg gacttgattt 4921 taattctgct gtagagtcgc aaccgggcat caaagacgca cgtcttttgg cctcggtttt 4981 ccagacgctg cgcgcatatt aaggaaagga acaatgacaa cattacttaa cccctatttt 5041 ggtgagtttg gcggcatgta cgtgccacaa atcctgatgc ctgctctgcg ccagctggaa 5101 gaagcttttg tcagtgcgca aaaagatcct gaatttcagg ctcagttcaa cgacctgctg 5161 aaaaactatg ccgggcgtcc aaccgcgctg accaaatgcc agaacattac agccgggacg 5221 aacaccacgc tgtatctcaa gcgtgaagat ttgctgcacg gcggcgcgca taaaactaac 5281 caggtgctgg ggcaggcgtt gctggcgaag cggatgggta aaaccgaaat catcgccgaa 5341 accggtgccg gtcagcatgg cgtggcgtcg gccctggcca gcgccctgct cggcctgaaa 5401 tgccgtattt atatgggtgc caaagacgtt gaacgccagt cgcctaacgt ttttcgtatg 5461 cgcttaatgg gtgcggaagt gatcccggtg catagcggtt ccgcgacgct gaaagatgcc 5521 tgtaacgagg cgctgcgcga ctggtccggt agttacgaaa ccgcgcacta tatgctgggc 5581 accgcagctg gcccgcatcc ttatccgacc attgtgcgtg agtttcagcg gatgattggc 5641 gaagaaacca aagcgcagat tctggaaaga gaaggtcgcc tgccggatgc cgttatcgcc 5701 tgtgttggcg gcggttcgaa tgccatcggc atgtttgctg atttcatcaa tgaaaccaac 5761 gtcggcctga ttggtgtgga gccaggtggt cacggtatcg aaactggcga gcacggcgca 5821 ccgctaaaac atggtcgcgt gggtatctat ttcggtatga aagcgccgat gatgcaaacc 5881 gaagacgggc agattgaaga atcttactcc atctccgccg gactggattt cccgtctgtc 5941 ggcccacaac acgcgtatct taacagcact ggacgcgctg attacgtgtc tattaccgat 6001 gatgaagccc ttgaagcctt caaaacgctg tgcctgcacg aagggatcat cccggcgctg 6061 gaatcctccc acgccttggc ccatgcgttg aaaatgatgc gcgaaaaccc ggataaagag 6121 cagctactgg tggttaacct ttccggtcgc ggcgataaag acatcttcac cgttcacgat 6181 attttgaaag cacgagggga aatctgatgg aacgctacga atctctgttt gcccagttga 6241 aggagcgcaa agaaggcgca ttcgttcctt tcgtcacgct cggtgatccg ggcattgagc 6301 agtcattgaa aattatcgat acgctaattg aagccggtgc tgacgcgctg gagttaggta 6361 tccccttctc cgacccactg gcggatggcc cgacgattca aaacgccact ctgcgcgcct 6421 ttgcggcagg tgtgactccg gcacaatgtt ttgaaatgct ggcactgatt cgccagaaac 6481 acccgaccat tcccattggc ctgttgatgt atgccaatct ggtgtttaac aaaggcattg 6541 atgagtttta tgcccagtgc gaaaaagtcg gcgtcgattc ggtgctggtt gccgatgtgc 6601 cagttgaaga gtccgcgccc ttccgccagg ccgcgttgcg tcacaacgtc gcacctatct 6661 tcatctgccc gccaaatgcc gatgacgacc tgctgcgcca gatagcctct tacggtcgtg 6721 gttacaccta tttgctgtca cgagcaggcg tgaccggcgc agaaaaccgc gccgcgttac 6781 ccctcaatca tctggttgcg aagctgaaag agtacaacgc tgcacctcca ttgcagggat 6841 ttggtatttc cgccccggat caggtaaaag cagcgattga tgcaggagct gcgggcgcga 6901 tttctggttc ggccattgtt aaaatcatcg agcaacatat taatgagcca gagaaaatgc 6961 tggcggcact gaaagttttt gtacaaccga tgaaagcggc gacgcgcagt taatcccaca 7021 gccgccagtt ccgctggcgg cattttaact ttctttaatg aagccggaaa aatcctaaat 7081 tcatttaata tttatctttt taccgtttcg cttaccccgg tcgatcgtyr acttacgtca 7141 tttttccgcc caacagtaat ataaacaaac aaattaaacc cgcaacataa caccagtaaa 7201 atcaataatt ttctctaagt cacttattcc tcaggtaatt cttaatatat ccagaatgtt 7261 cctcaaaata tattttccct ctatcttctc gttgcgctta atttgactaa ttctcattag 7321 cgactaattt taatgagtgt cgacacacaa cactcatatt aatgaaacaa tgcaacgcaa 7381 cgggagaaat aacatggccg aacatcgtgg tggttcagga aatttcgccg aagaccgtga 7441 gaaggcatcc gacgcagccg taaaggcggt cagcatagcg gcggtaattt taaaaatgat 7501 cgcaacgcgc atctgaagcg ggtaaaaaag gcggtyrac //