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ID PGEX4T1 preliminary; circular DNA; SYN; 4969 BP.
XX
AC IG1074; M21676; M97937;
XX
DT 01-FEB-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli plasmid vector pGEX-4T1 - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RC plasmid from pSj1 & pIC19H
RC pSj10 from plasmid & ptac or ptac12deltaEco
RC p4.5 from pSj5 & pMC9
RC pSj10Bam7Stop7 from pSj10
RC pGEX-1 from pBR322 & pSj10Bam7Stop7 & p4.5 & oligo
RC pGEX-2T from pGEX-1 & oligo, thrombin protease site
RC pGEX-3X from pGEX-1 & oligo, factor Xa site
RA Smith D.B., Johnson K.S.;
RT "Single-step purification of polypeptides expressed in Escherichia
RT coli as fusions with glutathione S-transferase";
RL Gene 67:31-40(1988).
XX
CC thrombin or factor Xa protease sites to cleave protein from fusion.
CC pGEX-1lambdaT, pGEX-4T-1, pGEX-5X-1 accept cDNA from lambda gt11 libs.
CC NM (pGEX-4T1)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (Pharmacia)
CC HO (E.coli)
CC CP ()
CC FN (expression)(cloning directional)
CC SE (affinity)(color yellow)
CC PA (pGEX-2T)
CC BR ()
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pGEX-4
FT 2. oligo, thrombin site
FT -> pGEX-4T-1 4969bp"
FT misc_binding 0..0
FT /note="MCS unique EcoRI-SmaI-SalI-XhoI-NotI"
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 unique PstI"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT CDS 0..0
FT /note="REP E. coli lacI repressor gene"
FT misc_binding 0..0
FT /note="SIT unique MluI"
FT misc_binding 0..0
FT /note="SIT unique BstEII"
FT misc_binding 0..0
FT /note="SIT unique ApaI"
FT misc_binding 0..0
FT /note="SIT unique BssHII"
FT misc_binding 0..0
FT /note="SIT unique EcoRV"
FT misc_binding 0..0
FT /note="SIT unique NarI"
FT promoter 0..0
FT /note="PRO E. coli tac (trp and lac)"
FT CDS 0..0
FT /note="GEN Schistosoma japonicum
FT glutathione S-transferase (GST)"
FT misc_binding 0..0
FT /note="SIT thrombin"
XX
SQ Sequence 4969 BP; 1225 A; 1202 C; 1292 G; 1250 T; 0 other;
acgttatcga 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 gttccgcgtg gatccccgga attcccgggt cgactcgagc
ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc
gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt
//