suter2@urz.unibas.ch (clemens suter-crazzolara) (05/01/91)
Hi there, we are looking for a simple pUC vector (preferably pUC 18 or 19) with a T7 promoter and a T3 promoter cloned in the polylinker. We already have pUC118, but this vector contains, as you know, #a origin for single stranded replication, and this hampers our cloning scheme. If you have this kind of vector, we would be very pleased to hear from you. Christian and Clemens
suter2@urz.unibas.ch (clemens suter-crazzolara) (05/13/91)
original message: > Hi there, > we are looking for a simple pUC vector (preferably pUC 18 > or 19) with a T7 promoter and a T3 promoter cloned in the > polylinker. > We already have pUC118, but this vector contains, as you know, > an origin for single stranded replication, and this hampers > our cloning scheme. > If you have this kind of vector, we would be very pleased to hear > from you. > Christian and Clemens this message is apparently very confusing, since we were recieved many answers (for which we are grateful) that we should use bluescript (or could it be that only few people know that this vector contains an ori for single stranded replication ?). But what we really want is a vector that looks like this: pUC sequences- T7 promoter-polylinker-T3 promoter-pUC sequences. Simple ? Yes ! M13 ori for ss replication ? No ! Before I start constructing myself, I would like to hear from anybody out there, whether he has this rather simple construct, or one like it. Clemens ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + + + Daar bij de haven, die mooie haven + + Waar je altijd horen kon: + + "We gaan aan wal !" + + + ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
BROE@AARDVARK.UCS.UOKNOR.EDU (Bruce Roe) (05/14/91)
You wrote: >> Hi there, >> we are looking for a simple pUC vector (preferably pUC 18 >> or 19) with a T7 promoter and a T3 promoter cloned in the >> polylinker. >> We already have pUC118, but this vector contains, as you know, >> an origin for single stranded replication, and this hampers >> our cloning scheme. >> If you have this kind of vector, we would be very pleased to hear >> from you. >> Christian and Clemens >this message is apparently very confusing, since we were recieved many >answers (for which we are grateful) that we should use bluescript (or >could it be that only few people know that this vector contains an ori >for single stranded replication ?). >But what we really want is a vector that looks like this: pUC sequences- >T7 promoter-polylinker-T3 promoter-pUC sequences. Simple ? Yes ! >M13 ori for ss replication ? No ! >Before I start constructing myself, I would like to hear from anybody out >there, whether he has this rather simple construct, or one like it. >Clemens Bethesda Research Labs. has to vectors called: pT7/T3a-18 and pT7/T3a-19. or at least they do according to their catalogue. They both are pUC bases and have either the pUC18 or 19 multiple cloning site flanked by the T7 and 73 promoters. Cheers, ****************************** Bruce A. Roe ****************************** * Professor of Chemistry and Biochemistry Dept. of Chem. and Biochem. * * INTERNET: BROE@aardvark.ucs.uoknor.edu University of Oklahoma * * BITNET: BROE@uokucsvx 620 Parrington Oval, Rm 208 * * AT&TNET: 405-325-4912 or 405-325-7610 Norman, Oklahoma 73019 * * FAXnet: 405-325-6111 * **************************************************************************** * ICBMnet: 35 deg 14 min North, 97 deg 27 min West * ****************************************************************************
NUM208JN@NRCCAD.NRC.CA (John Nash) (05/14/91)
G'day folk,
I saw this today...
*Date: Mon, 13 May 91 18:34 CDT
*From: Bruce Roe <BROE@aardvark.ucs.uoknor.edu>
*Subject: re: pUC vectors
*To: bio-soft@genbank.bio.net
Shouldn't this be "methods @genbank.bio.net"? Not a complaint from me,
just that it may get a more aappropriate audience there.
*X-Vms-To: IN%"bio-soft@genbank.bio.net"
*You wrote:
*>> Hi there,
*>> we are looking for a simple pUC vector (preferably pUC 18
*>> or 19) with a T7 promoter and a T3 promoter cloned in the
*>> polylinker.
*>> We already have pUC118, but this vector contains, as you know,
*>> an origin for single stranded replication, and this hampers
*>> our cloning scheme.
Shortened for brevity...
and
*>But what we really want is a vector that looks like this: pUC sequences-
*>T7 promoter-polylinker-T3 promoter-pUC sequences. Simple ? Yes !
*>M13 ori for ss replication ? No !
and
* Bethesda Research Labs. has to vectors called:
*pT7/T3a-18 and pT7/T3a-19. or at least they do according to their
*catalogue. They both are pUC bases and have either the pUC18 or 19
*multiple cloning site flanked by the T7 and 73 promoters.
I use the pT7/T3-alpha vectors, and I hate to report that they have
M13 (or f1) ss ori sites.
Sorry to be a bubble burster, but I think that Clemens may have to
start constructing ;-(
cheers,
John,
--------------------------------------------------------------
John H.E. Nash <Bitnet: NUM208JN@NRCCAD.NRC.CA >
Institute for Biological Sciences,
National Research Council of Canada,
Ottawa, Canada K1A 0R6.
Phone: (613) 990-0990 Fax: (613) 952-9092.
==============================================================
P.S. It's a BOY!TAO%OPUS@mcoiarc.bitnet (05/14/91)
I could not help giving my 2 cents worth regarding the posting on pUC vecter. It seems to me that people of the electronic age are even more narrowly-minded than their "ancestors". Maybe they are lazier too. Answer to the question is the pBS vector from Strategene. Even if one doesn't have the money to buy it, one could still do some simple restriction digestion and ligation to satisfy the experimental need [since the pUC based vector is available to the questioneer, with an unwanted feature, right?]. Problems tend to be unique for individual project, customerization usually would be the only way out. To be a little deligent and broad-minded will always help. Another thing is whoever answering the posting is actually doing the questioneer a favor. I hope that everyone could keep this in mind. Tao Tao "My name has nothing to do with the Graduate Student Taoism, even though I am a believer of it." Department of Microbiology Medical College of Ohio Toledo, Ohio 43699-0008
suter2@urz.unibas.ch (clemens suter-crazzolara) (05/16/91)
i think tao was pretty rude
suter2@urz.unibas.ch (clemens suter-crazzolara) (05/16/91)
to those netters that are interested................... the vectors pT7/T3-18 or -19 of Gibco BRL and.... the pT3t7 lac vectors of Boehringer...... do contain pUC sequences and the T7 and T3 promoters. They do NOT contain an ori for ss DNA replication, in other words, they are plasmids, not phagemids. The first one is only 2239 bp in size. I am sorry to have bothered you with so much bullshit about these vectors. Tao Tao is probably just as frustrated as you are........