LOCUS       sequence-128886-        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST tagged GIV-CT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  AUTHORS   Garcia-Marcos M, Ghosh P, Ear J, Farquhar MG
  TITLE     A structural determinant that renders G alpha(i) sensitive to 
            activation by GIV/girdin is required to promote cell migration.
  JOURNAL   J Biol Chem. 2010 Apr 23;285(17):12765-77. doi: 
            10.1074/jbc.M109.045161. Epub 2010 Feb 15.
  PUBMED    20157114
REFERENCE   2  (bases 1 to 5702)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M109.045161"; journalName: "J Biol Chem"; date: 
            "2010-04-23- 23"; volume: "285"; issue: "17"; pages: "12765-77"
COMMENT     Sequence Label: GST-GIV-CT
FEATURES             Location/Qualifiers
     source          1..5702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..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'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1752..1774)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1919..1937)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2005..2109
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2110..2178,2179..2970)
                     /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"
     primer_bind     complement(2328..2347)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3141..3729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3630..3649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3883..3900
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3973..4050
                     /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             4051..5133
                     /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"
     primer_bind     complement(4070..4089)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5146..5167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5182..5187,5188..5205,5206..5212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5220..5236
                     /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)."
     primer_bind     5244..5260
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5244..5260
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5256..5702,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5276..5293)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5276..5292)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5285..5307)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
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 gatccccgga attcggagaa tttagcctgc
      961 ttcatgacca tgaggcttgg tccagcagtg gtagcagtcc aatccagtac ttgaaaagac
     1021 agaccagatc aagcccagtg ctccagcaca aaatatctga aacactggag agtcgacatc
     1081 acaagatcaa aactggttcc cctggaagtg aagttgttac tctacaacag tttttggaag
     1141 aaagcaataa gcttacctca gtacagataa agtcctcaag tcaagagaat cttttagatg
     1201 aagtaatgaa aagtttgtct gtctcttctg actttttggg aaaagacaaa ccagttagct
     1261 gtggtctggc caggtcagta agtggaaaaa ccccagggga cttctatgat agacggacaa
     1321 ctaagcctga gtttttgaga cctggtcctc gaaaaactga agatacctac ttcattagtt
     1381 ctgcgggaaa acctacacca ggcactcaag gaaaaataaa attagtaaaa gaatcttctc
     1441 tgtcacgaca atcaaaagat agtaaccctt atgcaacttt acctcgtgca agcagcgtga
     1501 tctcaactgc cgaaggaact acacgaagga caagcatcca tgattttttg accaaggaca
     1561 gtagactgcc tatatcagtt gattcaccac cagctgctgc tgacagcaac accactgcag
     1621 catctaatgt ggacaaagta caagaaagca gaaattcaaa aagcaggtct agggagcaac
     1681 aaagctccta agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat
     1741 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     1801 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     1861 gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc
     1921 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     1981 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2041 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2101 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2161 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2221 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2281 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2341 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2401 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2461 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2521 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2581 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2641 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2701 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2761 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2821 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2881 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2941 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3001 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3061 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3121 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3181 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3241 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3301 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3361 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3421 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3481 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3541 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3601 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3661 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3721 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3781 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3841 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3901 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3961 cgcataaatt ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
     4021 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
     4081 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     4141 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     4201 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     4261 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     4321 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     4381 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     4441 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     4501 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     4561 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     4621 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     4681 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     4741 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     4801 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     4861 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     4921 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     4981 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     5041 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     5101 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     5161 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     5221 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg
     5281 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5341 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5401 cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag
     5461 cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct
     5521 caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtaacc tatcccatta
     5581 cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
     5641 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa
     5701 tt
//
