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
//