ID PGEX2TK preliminary; circular DNA; SYN; 4969 BP. XX AC IG1072; 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-2TK - 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 pGEX-2TK can label expressed protein in vitro by site for cAMP- CC dependent protein kinase from heart muscle. 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-2TK) 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 (p4.5)(pS) CC BR () CC OF () CC OR () XX FH Key Location/Qualifiers FH FT misc_feature 0..0 FT /note="1. pGEX-2T 4948bp FT 2. Schistosoma japonicum, GST gene FT -> pGEX-2TK 4969bp" FT misc_binding 0..0 FT /note="MCS unique BamHI-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 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; 1198 C; 1290 G; 1256 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 gatctcgtcg tgcatctgtt ggatccccgg gaattcatcg 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 //