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ID   PGEXKN     preliminary; circular DNA; SYN; 4966 BP.
XX
AC   IG5003; ATCC77332;
XX
DT   01-OCT-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pGEX-KN - incomplete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-45
RC   pGEX-KN, pGEX-KT from pGEX-1
RA   Hakes D.J., Dixon J.E.;
RT   "New vectors for high level expression of recombinant proteins in
RT   bacteria";
RL   Anal. Biochem. 202:293-298(1992).
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CC   Deposited by: David J. Hakes, Jack E. Dixon
CC   The glutathione S-transferase (GST) fusion protein can be purified by
CC   glutathione affinity chromatography, and the desired polypeptide
CC   released from the fusion product by thrombin. [1]
CC   Expression vector for rapid purification of fusion proteins that
CC   contain no amino terminal extensions after thrombin cleavage. The
CC   amino acid after the initiator methionine must be charged. [1]
CC   Cloning into this vector requires amplification of the gene using
CC   oligonucleotides prepared as in the reference and encoding the first 4
CC   amino acids of a thrombin recognition sequence. [1]
CC   Constructed from pGEX-1 by inserting an oligonucleotide at the BamHI
CC   site which encodes the glycine kinker and a NotI site. [1]
CC   The order of the major features in this plasmid is:  pMB1 ori - lacIV
CC   - lacZ - tac promoter - GST - glycine kinker - NotI/MCS/EcoRI -
CC   ampR. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   NotI--5.0; EcoRI/PstI--4.0, 1.0; BamHI/EcoRV--3.2, 1.8. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pGEX-KN)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli)(E.coli HB101)(E.coli JM101)(E.coli TG1)(E.coli BL21)
CC   CP ()
CC   FN (cloning)(expression)(affinity)
CC   SE ()
CC   PA (pGEX-1)
CC   BR (pGEX-KT)(pGEX-KG)
CC   OF (pGEX-KN-PTPU323)
CC   OF (pGEX-KN-LAMBDA-PP)
CC   OF (pGEX-KN-YERSINIA-PP)
CC   OR (yes)
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FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pGEX-1 BamHI 4941bp
FT                   2. oligo BamHI-BamHI 25bp, glycine
FT                   -> pGEX-KN 4966bp"
FT   promoter        0..0
FT                   /note="PRO E. coli tac promoter"
FT   CDS             0..0
FT                   /note="GEN Schistosoma japonicum
FT                   glutathione S-transferase (GST)"
FT   misc_feature    0..0
FT                   /note="oligonucleotide insert into pGEX-1 at BamHI"
FT   misc_feature    0..0
FT                   /note="SIT glycine kinker"
FT   misc_binding    0..0
FT                   /note="MCS unique NotI-BamHI-SmaI-EcoRI"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   misc_binding    0..0
FT                   /note="SIT PstI"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="REP E. coli lacIq repressor gene"
FT   misc_binding    0..0
FT                   /note="SIT EcoRV"
FT   CDS             0..0
FT                   /note="GEN E. coli beta-galactosidase gene (lacZ)"
XX
SQ   Sequence 4966 BP; 1223 A; 1199 C; 1295 G; 1249 T; 0 other;
     agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
     gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
     aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
     gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
     ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
     tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
     ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
     ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
     atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
     tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
     aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
     ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
     atcctccaaa atcggatctg ggtggtggtg gcggccgcgg atccccggga attcatcgtg
     actgactgac gatctgcctc gcgcgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat
     ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca
     ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg
     tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg
     gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca
     cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca atccgccgtt
     tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg
     gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaatt
//