LOCUS       Exported                5809 bp ds-DNA     circular SYN 07-OCT-2019
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 Doc2B(125) C2AB WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5809)
  AUTHORS   GE Healthcare
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 8, 2019 from SnapGene Viewer 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5809
                     /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"
     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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             936..947
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=Linker
                     /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)."
                     /translation="GSGS"
     CDS             948..1799
                     /codon_start=1
                     /label=rDoc2b C2AB
                     /translation="TALGTLDFSLLYDQENNALHCTISKAKGLKPMDHNGLADPYVKLH
                     LLPGASKANKLRTKTLRNTLNPSWNETLTYYGITDEDMIRKTLRISVCDEDKFRHNEFI
                     GETRVPLKKLKPNHTKTFSICLEKQLPVDKAEDKSLEERGRILISLKYSSQKQGLLVGI
                     VRCAHLAAMDANGYSDPYVKTYLKPDVDKKSKHKTAVKKKTLNPEFNEEFCYEIKHGDL
                     AKKTLEVTVWDYDIGKSNDFIGGVVLGINAKGERLKHWFDCLKNKDKRIERWHTLTNEL
                     PG"
     misc_feature    1788
                     /label=A->C ILE->LEU
     misc_feature    1797..1799
                     /label=C-term deletion
     misc_feature    1811..1821
                     /label=stop codons
                     /note="stop codons in all three reading frames"
     promoter        2112..2216
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2217..3077
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3248..3836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4080..4157
                     /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."
     CDS             4158..5240
                     /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)."
                     /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 gatccggatc tggatctacc gccctgggta
      961 cactggactt cagtctgctc tatgaccagg agaacaacgc actgcactgc accatcagca
     1021 aagccaaggg cctgaagccg atggatcaca atggactggc tgatccctac gtcaaactac
     1081 acttgctgcc tggagccagc aaggcaaata agctcagaac aaaaacccta cggaacactc
     1141 tgaacccctc ctggaacgag accctcacct attacgggat cacggatgag gacatgatcc
     1201 gaaagaccct gaggatctcc gtgtgtgacg aggacaaatt ccgccacaac gagttcatcg
     1261 gagagactcg agtgcccctg aagaagctga aacccaacca caccaagaca ttcagcatct
     1321 gcctggagaa gcagctgccg gtggacaagg cagaggacaa gtccctggag gagcgtggcc
     1381 gcatcctcat ctctctcaag tacagctcac agaagcaggg cctgctggtg ggcatcgttc
     1441 gctgcgcaca cctggcggcc atggatgcta acggctactc agacccctac gtgaaaacat
     1501 atctgaaacc agatgtagac aagaaatcca agcataagac cgcagtcaag aagaaaacac
     1561 taaaccctga attcaatgag gagttctgtt acgagatcaa gcatggagac ctagccaaaa
     1621 agaccctgga ggtcactgtc tgggactatg atattggaaa atccaatgat ttcatcggtg
     1681 gggtggttct gggcatcaat gccaagggtg agcgcctgaa gcactggttt gactgtctga
     1741 agaacaagga caagaggatt gagcgttggc acacgctcac caatgagctc ccagggtagc
     1801 ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     1861 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     1921 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     1981 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
     2041 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2101 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2161 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2221 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2281 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2341 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2401 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     2461 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2521 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2581 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2641 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2701 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2761 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2821 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2881 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2941 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3001 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3061 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3121 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3181 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3241 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3301 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3361 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3421 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3481 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3541 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3601 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3661 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3721 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3781 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3841 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3901 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3961 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4021 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
     4081 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     4141 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     4201 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     4261 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     4321 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     4381 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     4441 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     4501 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     4561 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     4621 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     4681 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     4741 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     4801 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     4861 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     4921 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     4981 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     5041 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     5101 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     5161 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5221 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5281 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5341 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
     5401 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5461 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5521 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
     5581 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
     5641 gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc
     5701 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     5761 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt
//
