LOCUS       Exported                7427 bp DNA     circular SYN 17-MAY-2019
DEFINITION  Bacterial vector for expressing GST fusion proteins with a 
            PreScission protease site. For other reading frames, use pGEX-6P-2 
            or pGEX-6P-3.
ACCESSION   U78872
VERSION     .
KEYWORDS    pGEX-6P-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7427)
  AUTHORS   GE Healthcare
  TITLE     Direct Submission
  JOURNAL   Exported Jun 14, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7427
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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     869..891
                     /label=pGEX 5' Sequencing Primer
                     /note="color: black; sequence: GGGCTGGCAAGCCACGTTTGGTG"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=PreScission site
                     /note="color: #cc99b2
                     Cleavage site after base 935"
                     /translation="LEVLFQGP"
     misc_feature    3429..3439
                     /label=stop codons
                     /note="stop codons in all three reading frames"
                     /note="color: #ff0000"
     primer_bind     complement(3477..3499)
                     /label=pGEX 3' Sequencing Primer
                     /note="color: black; sequence: CCGGGAGCTGCATGTGTCAGAGG"
     promoter        3730..3834
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3835..4695
                     /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: 3835 .. 3903 / #ccffcc / signal sequence
                        2: 3904 .. 4695 / #ccffcc
                     Cleavage site after base 3903"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4866..5454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     promoter        5698..5775
                     /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             5776..6858
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     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 gaagttctgt tccaggggcc cgtacgcgcc ggctccatgg
      961 cgaccgggct cggggagccg gtctatggac tttccgaaga cgagggagag tcccgtattc
     1021 tcagagtaaa agttgtttct ggaattgatc tcgccaaaaa ggacatcttt ggagccagtg
     1081 atccgtatgt gaaactttca ttgtacgtag cggatgagaa tagagaactt gctttggtcc
     1141 agacaaaaac aattaaaaag acactgaacc caaaatggaa tgaagaattt tatttcaggg
     1201 taaacccatc taatcacaga ctcctatttg aagtatttga cgaaaataga ctgacacgag
     1261 acgacttcct gggccaggtg gacgtgcccc ttagtcacct tccgacagaa gatccaacca
     1321 tggagcgacc ctatacattt aaggactttc tcctcagacc aagaagtcat aagtctcgag
     1381 ttaagggatt tttgcgattg aaaatggcct atatgccaaa aaatggaggt caagatgaag
     1441 aaaacagtga ccagagggat gacatggagc atggatggga agttgttgac tcaaatgact
     1501 cggcttctca gcaccaagag gaacttcctc ctcctcctct gcctcccggg tgggaagaaa
     1561 aagtggacaa tttaggccga acttactatg tcaaccacaa caaccggacc actcagtggc
     1621 acagaccaag cctgactacg ccgggtctgc cttcaggctg ggaagaaaga aaagatgcta
     1681 aggggcgcac atactatgtc aatcataaca atcgaaccac aacttggact cgacctatca
     1741 tgcagcttgc agaagatggt gcgtccggat cagccacaaa cagtaacaac catctaatcg
     1801 agcctcagat ccgccggcct cgtagcctca gctcgccaac agtaacttta tctgccccgc
     1861 tggagggtgc caaggactca cccgtacgtc gggctgtgaa agacaccctt tccaacccac
     1921 agtccccaca gccatcacct tacaactccc ccaaaccaca acacaaagtc acacagagct
     1981 tcttgccacc cggctgggaa atgaggatag cgccaaacgg ccggcccttc ttcattgatc
     2041 ataacacaaa gactacaacc tgggaagatc cacgtttgaa atttccagta catatgcggt
     2101 caaagacatc tttaaacccc aatgaccttg gcccccttcc tcctggctgg gaagaaagaa
     2161 ttcacttgga tggccgaacg ttttatattg atcataatag caaaattact cagtgggaag
     2221 acccaagact gcagaaccca gctattactg gtccggctgt cccttactcc agagaattta
     2281 agcagaaata tgactacttc aggaagaaat taaagaaacc tgctgatatc cccaataggt
     2341 ttgaaatgaa acttcacaga aataacatat ttgaagagtc ctatcggaga attatgtccg
     2401 tgaaaagacc agatgtccta aaagctagac tgtggattga gtttgaatca gagaaaggtc
     2461 ttgactatgg gggtgtggcc agagaatggt tcttcttact gtccaaagag atgttcaacc
     2521 cctactacgg cctctttgag tactctgcca cggacaacta cacccttcag atcaacccta
     2581 attcaggcct ctgtaatgag gatcatttgt cctacttcac ttttattgga agagttgctg
     2641 gtctggccgt atttcatggg aagctcttag atggtttctt cattagacca ttttacaaga
     2701 tgatgttggg aaagcagata accctgaatg acatggaatc tgtggatagt gaatattaca
     2761 actctttgaa atggatcctg gagaatgacc ctactgagct ggacctcatg ttctgcatag
     2821 acgaagaaaa ctttggacag acatatcaag tggatttgaa gcccaatggg tcagaaataa
     2881 tggtcacaaa tgaaaacaaa agggaatata tcgacttagt catccagtgg agatttgtga
     2941 acagggtcca gaagcagatg aacgccttct tggagggatt cacagaacta cttcctattg
     3001 atttgattaa aatttttgat gaaaatgagc tggagttgct catgtgcggc ctcggtgatg
     3061 tggatgtgaa tgactggaga cagcattcta tttacaagaa cggctactgc ccaaaccacc
     3121 ccgtcattca gtggttctgg aaggctgtgc tactcatgga cgccgaaaag cgtatccggt
     3181 tactgcagtt tgtcacaggg acatcgcgag tacctatgaa tggatttgcc gaactttatg
     3241 gttccaatgg tcctcagcta tttacaatag agcaatgggg cagtcctgag aaactgccca
     3301 gagctcacac atgctttaat cgccttgact tacctccata tgaaaccttt gaagatttac
     3361 gagagaaact tctcatggcc gtggaaaatg ctcaaggatt tgaaggggtg gattaagcgg
     3421 ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3481 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3541 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     3601 tcacgtagcg atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta
     3661 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3721 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3781 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3841 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3901 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3961 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4021 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     4081 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4141 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4201 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4261 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4321 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     4381 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4441 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4501 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4561 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4621 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4681 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4741 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4801 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4861 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4921 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4981 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5041 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5101 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5161 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5221 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5281 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5341 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5401 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5461 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5521 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5581 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5641 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac
     5701 accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     5761 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     5821 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     5881 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     5941 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     6001 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     6061 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     6121 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     6181 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     6241 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     6301 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     6361 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     6421 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     6481 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     6541 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     6601 accgaagaca gctcatgtta tatcccgccg tcaaccacca tcaaacagga ttttcgcctg
     6661 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     6721 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     6781 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6841 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6901 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6961 atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac
     7021 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     7081 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     7141 gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct
     7201 ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc
     7261 acggttacga tgcgcccatc tacaccaacg taacctatcc cattacggtc aatccgccgt
     7321 ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct
     7381 ggctacagga aggccagacg cgaattattt ttgatggcgt tggaatt
//
