LOCUS       Exported                5907 bp DNA     circular SYN 15-OCT-2017
DEFINITION  Bacterial vector for expressing fusion proteins with a thrombin 
            site. For other reading frames, use pGEX-4T-2 or pGEX-4T-3.
ACCESSION   U13853
VERSION     .
KEYWORDS    pGEX-4T-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5907)
  AUTHORS   GE Healthcare
  TITLE     Direct Submission
  JOURNAL   Exported Nov 16, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5907
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     misc_feature    183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
                     /note="This forward directional feature has 3 segments:
                        1: 183 .. 188 / #ffffff / -35
                        2: 189 .. 204 / #ffffff
                        3: 205 .. 211 / #ffffff / -10"
     protein_bind    219..235
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /note="color: #cc99b2"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     600..620
                     /label=GST-115F-Primer
                     /note="color: black; sequence: TTTGAAACTCTCAAAGTTGAT"
     primer_bind     869..891
                     /label=GST detect F
                     /note="color: black; sequence: GGGCTGGCAAGCCACGTTTGGTG"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /note="color: #cc99b2
                     Cleavage site after base 929"
                     /translation="LVPRGS"
     misc_feature    919..938
                     /label=?????
                     /note="color: #a6acb3"
     misc_feature    930..938
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     misc_feature    939..1904
                     /label=FBL
                     /note="color: #a6acb3"
     misc_feature    1905..1924
                     /label=?????
                     /note="color: #a6acb3"
     misc_feature    1909..1919
                     /label=stop codons
                     /note="stop codons in all three reading frames"
                     /note="color: #ff0000"
     primer_bind     complement(1957..1979)
                     /label=GST detect R
                     /note="color: black; sequence: CCGGGAGCTGCATGTGTCAGAGG"
     promoter        2210..2314
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2315..3175
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 2315 .. 2383 / #ccffcc / signal sequence
                        2: 2384 .. 3175 / #ccffcc
                     Cleavage site after base 2383"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3346..3934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     promoter        4178..4255
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4256..5338
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatccccgat gaagccagga ttcagtcccc
      961 gtgggggtgg ctttggcggc cgagggggct ttggtgaccg tggtggtcgt ggaggccgag
     1021 ggggctttgg cgggggccga ggtcgaggcg gaggctttag aggtcgtgga cgaggaggag
     1081 gtggaggcgg cggcggcggt ggaggaggag gaagaggtgg tggaggcttc cattctggtg
     1141 gcaaccgggg tcgtggtcgg ggaggaaaaa gaggaaacca gtcggggaag aatgtgatgg
     1201 tggagccgca tcggcatgag ggtgtcttca tttgtcgagg aaaggaagat gcactggtca
     1261 ccaagaacct ggtccctggg gaatcagttt atggagagaa gagagtctcg atttcggaag
     1321 gagatgacaa aattgagtac cgagcctgga accccttccg ctccaagcta gcagcagcaa
     1381 tcctgggtgg tgtggaccag atccacatca aaccgggggc taaggttctc tacctcgggg
     1441 ctgcctcggg caccacggtc tcccatgtct ctgacatcgt tggtccggat ggtctagtct
     1501 atgcagtcga gttctcccac cgctctggcc gtgacctcat taacttggcc aagaagagga
     1561 ccaacatcat tcctgtgatc gaggatgctc gacacccaca caaataccgc atgctcatcg
     1621 caatggtgga tgtgatcttt gctgatgtgg cccagccaga ccagacccgg attgtggccc
     1681 tgaatgccca caccttcctg cgtaatggag gacactttgt gatttccatt aaggccaact
     1741 gcattgactc cacagcctca gccgaggccg tgtttgcctc cgaagtgaaa aagatgcaac
     1801 aggagaacat gaagccgcag gagcagttga cccttgagcc atatgaaaga gaccatgccg
     1861 tggtcgtggg agtgtacagg ccacccccca aggtgaagaa ctgaatcgtg actgactgac
     1921 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     1981 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2041 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     2101 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2161 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2221 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2281 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2341 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2401 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2461 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2521 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     2581 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2641 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2701 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2761 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2821 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     2881 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2941 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3001 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3061 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3121 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3181 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3241 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3301 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3361 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3421 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3481 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3541 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3601 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3661 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3721 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3781 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3841 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3901 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3961 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4021 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4081 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4141 tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat ggtgcaaaac
     4201 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     4261 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     4321 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     4381 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     4441 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     4501 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     4561 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     4621 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     4681 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     4741 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     4801 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     4861 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     4921 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     4981 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     5041 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     5101 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     5161 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     5221 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     5281 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5341 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5401 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5461 atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5521 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5581 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     5641 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     5701 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     5761 tacaccaacg taacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     5821 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     5881 cgaattattt ttgatggcgt tggaatt
//
