plasmid-sequence of YIplac204
Francis Ouellette
francis at borduas.nlm.nih.gov
Thu Jan 13 14:17:25 EST 1994
HANNES at bch.univie.ac.at (Hannes Klump) writes:
: Hi everybody! Has anybody of you got the nucleotide-sequence of
: the 3.54kb yeast integration vector "YIplac204" ? I am afraid I
: haven't got further information about the vector (who constructed it,
: etc.).
: Thanks a lot in advance, Hannes.
Dear Hannes,
This one (as many others :) can be retrived from the retrieve server
at NCBI. Mail to:
retrieve at ncbi.nlm.nih.gov
with this message:
datalib genbank
begin
YIplac204
To know more about this Email server, send help to the same address,
and you will get a detailed HELP document on how to use it.
A current list of all the yeast vectors present in GenBank has been
put together by David Stillman, and can be obtained by anonymous FTP
at ncbi.nlm.nih.gov in the /repository/yeast directory as:
yeast.vec
(login as anonymous and use your email address as your password)
regards,
francis
--
| B.F. Francis Ouellette
|
| francis at ncbi.nlm.nih.gov
Date: Thu, 13 Jan 1994 14:07:14 +0500
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=============================================================================
To Obtain Help Documentation: send e-mail to 'retrieve at ncbi.nlm.nih.gov'
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Database: GenBank Updates (80.0+, 01/13/94)
Query: yiplac204
Parse status: OK: 1 document retrieved.
Documents selected: 1-1 (up to 1000 lines)
LOCUS CVYIPL204 3545 bp DNA SYN 08-DEC-1993
DEFINITION Saccharomyces cerevisiae/E. coli shuttle vector, YIplac204
ACCESSION X75461 L26357
KEYWORDS TRP1 gene.
SOURCE Cloning vector
ORGANISM Cloning vector
Artificial sequences; Cloning vectors.
REFERENCE 1 (bases 1 to 3545)
AUTHORS Gietz,R.D.
TITLE Direct Submission
JOURNAL Submitted (13-MAY-1993) GIETZ at BLDGHSC.LAN1.UMANITOBA.CA.
STANDARD full automatic
REFERENCE 2 (bases 1 to 3545)
AUTHORS Gietz,R.D. and Sugino,A.
TITLE New yeast-Escherichia coli shuttle vectors constructed with in
vitro mutagenized yeast genes lacking six-base pair restriction
sites
JOURNAL Gene 74, 527-534 (1988)
STANDARD full automatic
COMMENT
Information taken from figure 2 , citation [2].
All constructions were initially made in the E. coli plasmid pBR322
for ease of manipulation. The yeast genes were inserted into
either
the EcoRI site or the Cla I site using PolIK and dNTPs to fill in
overhangs and T4 DNA ligase for blunt-end ligation. Any
restriction
site regenerated by this process was destroyed following digestion
with the appropriate restriction enzyme using PolIK and dNTPs to
fill
in overhangs and T4 DNA ligase to ligate blunt ends. Once the
constructions had been verified, the yeast genes were moved into
the
plasmid vector pUC19. To do this, the yeast genes were excised from
the pBR322 DNA backbone by HindIII digestion followed by mung-bean
exonuclease treatment to produce blunt ends. Then the DNA was
digested by AatII, the fragment containing yeast DNA was isolated
from agarose gels and ligaed to the large AatII-Nde I fragment of
pUC19 which had the NdeI end filled in with PolIk. The YCplac
vectors
(ARS1-CEN4) were constructed by blunt-end ligation of the 1.4kb Eco
RI
fragment of the TRP1 ARS1 gene(Struhl et al., 1979) into the EcoRI
site of pBR322 (pT322) and subsequent blunt-end ligation of the
functional 850-bp PvuII-HpaI CEN4 fragment (Mann and Davis, 1986)
into
the ClaI site of pT322(pTC1). This construction was then moved
into
pUC19 as described above to produce YCplac22. The other YCplac
vectors were made by blunt-end ligation of the EcoRV fragment from
pTC1, containing the ARS1 and CEN4 sequences, into the ClaI cite
of
pBR322 (pAC5). The 1.1-kb HindIII-SmaI fragment of the URA3 gene
or
the 1.6-kb HpaI-ACC I fragment containing LEU2 was inserted at
the
EcoRI site of pAC5 as described above. These constructs were also
moved into the plasmid pUC19 as described above givingrise to
YCplac33
and YCplac111, respectively. The YEplac vetor series (2mu origin)
were constructed by blunt-end ligation of the 1.5-kb Sau3A
fragment
from YEp24 which contains the 2 mu m ori (Broach, 1983), into the
ClaI site of pBR322 to produce p2mu19. The XbaI site of this
fragment was removed by filling in with PolIk after digestion and
blunt-end religation of this site. The three yeast genes were
ligated into the EcoRI site of this plasmid. The fragments used
for
the LEU2 and the URA3 constructs were the same as those used in the
YCplac constructions, however, the YEplac112 vector (TRPI) contains
the 850-bp EcoRI-BglII fragment of the TRP1 gene which does not
contain
ARS1 function. The YIplac vectors were constructed by ligating each
yeast gene fragment used in YEplac vector constructions into the
EcoO109 site of the plasmid pUC19 using PolIk to fill the sticky
ends.
FEATURES Location/Qualifiers
source 1..3545
/organism="Cloning vector"
BASE COUNT 899 a 906 c 817 g 923 t
ORIGIN
1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
481 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
541 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
601 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
661 caccgtcatc accgaaacgc gcgagacgaa agggcgatct tttatgcttg cttttcaaaa
721 ggcttgcagg caagtgcaca aacaatactt aaataaatac tactcagtaa taacctattt
781 cttagcattt ttgacgaaat ttgctatttt gttagagtct tttacaccat ttgtctccac
841 acctccgctt acatcaacac caataacgcc atttaatcta agcgcatcac caacattttc
901 tggcgtcagt ccaccagcta acataaaatg taagctctcg gggctctctt gccttccaac
961 ccagtcagaa atcgagttcc aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa
1021 caagggaata aacgaatgag gtttctgtga agctgcactg agtagtatgt tgcagtcttt
1081 tggaaatacg agtcttttaa taactggcaa accgaggaac tcttggtatt cttgccacga
1141 ctcatctcca tgcagttgga cgatatcaat gccgtaatca ttgaccagag ccaaaacatc
1201 ctccttaggt tgattacgaa acacgccaac caagtatttc ggagtgcctg aactattttt
1261 atatgctttt acaagacttg aaattttcct tgcaataacc gggtcaattg ttctctttct
1321 attgggcaca catataatac ccagcaagtc agcatcggaa tctagtgcac attctgcggc
1381 ctctgtgctc tgcaagccgc aaactttcac caatggacca gaactacctg tgaaattaat
1441 aacagacata ctccaagctg cctttgtgtg cttaatcacg tatactcacg tgctcaatag
1501 tcaccaatgc cctccctctt ggccctctcc ttttcttttt tcgaccgaat tggcctcgtg
1561 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
1621 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
1681 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
1741 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
1801 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
1861 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
1921 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
1981 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
2041 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
2101 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
2161 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
2221 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
2281 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
2341 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
2401 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
2461 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
2521 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
2581 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
2641 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
2701 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
2761 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
2821 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
2881 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
2941 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
3001 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
3061 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
3121 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
3181 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
3241 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
3301 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
3361 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
3421 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
3481 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
3541 gaaga
//
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