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ID   PGEXKT     preliminary; circular DNA; SYN; 4978 BP.
XX
AC   IG5002; ATCC77331;
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-KT - complete.
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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   1-57
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 and
CC   release of proteins with fewer extraneous N-terminal amino acids. [1]
CC   The resulting protein contains two additional amino acids at the amino
CC   terminus (Gly-Ser). [1]
CC   Constructed from pGEX-1 by inserting an oligonucleotide at the BamHI
CC   site which encodes the glycine kinker and thrombin recognition site
CC   to enhance cleavage of the fusion protein. [1]
CC   The order of the major features in the plasmid is:  pMB1 ori - lacIq -
CC   lacZ - tac promoter - GST - glycine kinker - thrombin cleavage site
CC   - BamHI/MCS/EcoRI - ampR. [1]
CC   Restriction digests of the clone give the following sizes (kb):
CC   BamHI--5.0; EcoRI/PstI--4.0, 1.0; EcoRI/EcoRV--3.2, 1.8. (ATCC staff)
CC   Medium is 1227 LB plus ampicillin.
CC   NM (pGEX-KT)
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-KN)(pGEX-KG)
CC   OF (pGEX-KT-PTPU323)
CC   OF (pGEX-KT-LAMBDA-PP)
CC   OF (pGEX-KT-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 37bp, glycine/thrombin site
FT                   -> pGEX-KT 4978bp"
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_feature    0..0
FT                   /note="SIT thrombin cleavage site"
FT   misc_binding    0..0
FT                   /note="MCS unique 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 4978 BP; 1223 A; 1200 C; 1298 G; 1257 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 tctggtggtg gtggtggtct ggttccgcgt ggatccccgg
     gaattcatcg tgactgactg acgatctgcc tcgcgcgcct cgcgcgtttc ggtgatgacg
     gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     ccatgaccca gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg
     tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     taaattccga caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
     atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt
     cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt
     gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa
     ctggcagatg cacggttacg atgcgcccat ctacaccaac gtaacctatc ccattacggt
     caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt
     tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaatt
//