LOCUS       Exported                5799 bp ds-DNA     circular SYN 13-DEC-2022
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 5799)
  AUTHORS   GE Healthcare
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 16, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5799
                     /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..938
                     /codon_start=1
                     /label=Start Codon
                     /translation="M"
     CDS             939..1790
                     /codon_start=1
                     /label=Syt9_Mus
                     /translation="SDKHQLGRLQYSLDYDFQTGQLLVGILQAQGLAALDLGGSSDPYV
                     SVYLLPDKRRRHETKVHRQTLNPHFGETFAFKVPYVELGGRVLVMAVYNFNRFSRNDAI
                     GEVRVPMSSVNLGRPVQAWRELQVAPKEEQEKLGDICFSLRYVPTAGKLTVIVLEAKNL
                     KKMDVGGLSDPYVKVHLLQGGKKVRKKKTTIKKNTLNPYYNEAFSFEVPCDQVQKVQVE
                     LTVLNYNKLGKNEAIGRVAVGAAVGGAGLRHWADMLANPRRPIAQWHSLRPPDRARPIP
                     AP"
     misc_feature    1791..1795
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1801..1811
                     /label=stop codons
                     /note="stop codons in all three reading frames"
     primer_bind     complement(1849..1871)
                     /label=pGEX 3' Sequencing Primer
     promoter        2102..2206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2207..3067
                     /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      3238..3826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4070..4147
                     /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             4148..5230
                     /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 gatccatgtc agacaaacac cagctaggcc
      961 gactgcagta ttcactggac tatgacttcc agactggcca gctcctggtg ggcatcttgc
     1021 aagctcaggg actggctgcg ttggacctag gaggttcctc agatccctat gttagtgtct
     1081 atctgctgcc agacaagcgg aggcgacatg agaccaaggt gcatcggcag accctgaatc
     1141 cacactttgg agagacattt gccttcaagg tcccatacgt ggaactaggg ggcagagtgc
     1201 tggtcatggc ggtgtataac tttaatcggt tctcccgcaa cgatgccatc ggggaggtgc
     1261 gggtgcctat gagttcagtg aacctggggc ggccggtgca ggcctggaga gagctgcagg
     1321 tggctcccaa agaggagcaa gagaaactgg gggacatctg cttctctctc cgttatgtcc
     1381 ccacggccgg gaagctcacc gtcattgtct tggaagctaa aaacctgaag aaaatggatg
     1441 taggaggact ctcagatccc tatgtgaagg tgcacctgct ccagggaggt aaaaaggttc
     1501 gcaagaagaa aaccaccatt aagaagaaca ccctgaaccc ctattacaac gaggccttca
     1561 gctttgaggt gccctgtgac caggtgcaga aagtccaggt ggagctgact gttctgaatt
     1621 ataacaaact ggggaagaat gaggccatcg ggagagtggc ggtgggcgca gcagtcggtg
     1681 gggctggcct gcggcactgg gctgacatgt tggccaaccc caggcgcccc attgcccagt
     1741 ggcactctct gcggccccct gacagagcca ggccaatacc tgcaccctga gaattcatcg
     1801 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     1981 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     2041 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2101 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2161 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2221 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2281 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2341 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2401 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     2461 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2521 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2581 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2641 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2701 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     2761 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2821 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2881 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2941 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3001 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3061 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3121 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3181 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3241 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3301 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3361 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3421 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3481 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3541 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3601 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3661 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3721 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3781 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3841 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3901 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3961 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4021 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     4081 atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     4141 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     4201 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     4261 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     4321 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     4381 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     4441 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     4501 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     4561 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     4621 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     4681 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     4741 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     4801 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     4861 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     4921 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     4981 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     5041 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     5101 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     5161 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5221 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     5281 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5341 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     5401 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5461 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5521 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     5581 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     5641 gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc gtttgttccc
     5701 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     5761 gaaggccaga cgcgaattat ttttgatggc gttggaatt
//
