LOCUS       Exported                5822 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) G361C
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5822)
  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..5822
                     /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"
     primer_bind     869..891
                     /label=pGEX 5' Sequencing Primer
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             936..946
                     /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="GSG"
     CDS             948..1812
                     /codon_start=1
                     /label=rDoc2b_C2AB_cys-less
                     /translation="ALGTLDFSLLYDQENNALHATISKAKGLKPMDHNGLADPYVKLHL
                     LPGASKANKLRTKTLRNTLNPSWNETLTYYGITDEDMIRKTLRISVADEDKFRHNEFIG
                     ETRVPLKKLKPNHTKTFSIALEKQLPVDKAEDKSLEERGRILISLKYSSQKQGLLVGIV
                     RAAHLAAMDANGYSDPYVKTYLKPDVDKKSKHKTAVKKKTLNPEFNEEFAYEIKHGDLA
                     KKTLEVTVWDYDICKSNDFIGGVVLGINAKGERLKHWFDALKNKDKRIERWHTLTNEIP
                     GAVLSD"
     primer_bind     948..974
                     /label=Doc2b_reassemble_for
     misc_feature    1005..1007
                     /label=C -> A
     misc_feature    1221..1223
                     /label=C->A
     misc_feature    1317..1319
                     /label=C->A
     misc_feature    1440..1442
                     /label=C->A
     misc_feature    1584..1586
                     /label=C->A
     misc_feature    1653..1655
                     /label=G->C
     misc_feature    1731..1733
                     /label=C->A
     primer_bind     complement(1806..1823)
                     /label=Doc2b-pET_IF_rev
     misc_feature    1824..1834
                     /label=stop codons
                     /note="stop codons in all three reading frames"
     primer_bind     complement(1872..1894)
                     /label=pGEX 3' Sequencing Primer
     promoter        2125..2229
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2230..3090
                     /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      3261..3849
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4093..4170
                     /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             4171..5253
                     /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 tggatcagcc ctgggtacac
      961 tggacttcag tctgctctat gaccaggaga acaacgcact gcacgcgacc atcagcaaag
     1021 ccaagggcct gaagccgatg gatcacaatg gactggctga tccctacgtc aaactacact
     1081 tgctgcctgg agccagcaag gcaaataagc tcagaacaaa aaccctacgg aacactctga
     1141 acccctcctg gaacgagacc ctcacctatt acgggatcac ggatgaggac atgatccgaa
     1201 agaccctgag gatctccgtg gcggacgagg acaaattccg ccacaacgag ttcatcggag
     1261 agactcgagt gcccctgaag aagctgaaac ccaaccacac caagacattc agcatcgcgc
     1321 tggagaagca gctgccggtg gacaaggcag aggacaagtc cctggaggag cgtggccgca
     1381 tcctcatctc tctcaagtac agctcacaga agcagggcct gctggtgggc atcgttcgcg
     1441 cggcacacct ggcggccatg gatgctaacg gctactcaga cccctacgtg aaaacatatc
     1501 tgaaaccaga tgtagacaag aaatccaagc ataagaccgc agtcaagaag aaaacactaa
     1561 accctgaatt caatgaggag ttcgcgtacg agatcaagca tggagaccta gccaaaaaga
     1621 ccctggaggt cactgtctgg gactatgata tttgcaaatc caatgatttc atcggtgggg
     1681 tggttctggg catcaatgcc aagggtgagc gcctgaagca ctggtttgac gcgctgaaga
     1741 acaaggacaa gaggattgag cgttggcaca cgctcaccaa tgagatccca ggggctgtac
     1801 tcagcgactg agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat
     1861 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     1921 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     1981 gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc
     2041 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     2101 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2161 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2221 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2281 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2341 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2401 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2461 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2521 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2581 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2641 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2701 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2761 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2821 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2881 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2941 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3001 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3061 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3121 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3181 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3241 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3301 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3361 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3421 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3481 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3541 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3601 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3661 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3721 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3781 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3841 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3901 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3961 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4021 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4081 cgcataaatt ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
     4141 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     4201 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     4261 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     4321 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     4381 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     4441 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     4501 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     4561 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     4621 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     4681 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     4741 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     4801 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     4861 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     4921 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     4981 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     5041 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     5101 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     5161 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     5221 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     5281 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     5341 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg
     5401 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5461 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5521 cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag
     5581 cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct
     5641 caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtaacc tatcccatta
     5701 cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
     5761 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa
     5821 tt
//
