LOCUS       sequence-204865-        5492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5492)
  AUTHORS   Chinthalapudi K, Rangarajan ES, Patil DN, George EM, Brown DT, Izard
            T
  TITLE     Lipid binding promotes oligomerization and focal adhesion activity 
            of vinculin.
  JOURNAL   J Cell Biol. 2014 Dec 8;207(5):643-56. doi: 10.1083/jcb.201404128.
  PUBMED    25488920
REFERENCE   2  (bases 1 to 5492)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2014-12-8- 8"; volume: "207"; 
            issue: "5"; pages: "643-56"
COMMENT     Sequence Label: pGEX-6P1-Vinculin tail
FEATURES             Location/Qualifiers
     source          1..5492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     765..782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        855..932
                     /gene="lacI"
                     /label=lacI promoter
     CDS             933..2015
                     /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(952..971)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2064..2069,2070..2087,2088..2094)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2102..2118
                     /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     2126..2142
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2126..2142
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2138..2665
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2158..2175)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2158..2174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2167..2189)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2767..2772,2773..2788,2789..2795)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2803..2819
                     /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             2842..3495
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3453..3475
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3502..3525
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(4126..4148)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4293..4311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4379..4483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4484..4552,4553..5344)
                     /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(4702..4721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      781 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
      841 accgcataaa ttccgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
      901 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
      961 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     1021 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     1081 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     1141 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     1201 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     1261 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     1321 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     1381 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     1441 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     1501 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     1561 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     1621 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     1681 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     1741 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     1801 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     1861 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccacgc
     1921 tagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     1981 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     2041 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     2101 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact
     2161 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2221 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2281 ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga
     2341 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc
     2401 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat
     2461 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt
     2521 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg
     2581 aattagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
     2641 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc
     2701 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat
     2761 gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat
     2821 ttcacacagg aaacagtatt catgtcccct atactaggtt attggaaaat taagggcctt
     2881 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
     2941 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
     3001 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
     3061 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
     3121 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
     3181 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
     3241 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
     3301 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
     3361 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
     3421 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
     3481 gaccatcctc caaaatcgga tctggaagtt ctgttccagg ggcccctggg atccggggag
     3541 gtgattaacc agccaatgat gatggctgcc agacagctcc atgatgaagc tcgcaaatgg
     3601 tccagcaagg gcaatgacat cattgcagca gccaagcgca tggctctgct gatggctgag
     3661 atgtctcggc tggtaagagg gggcagtggt accaagcggg cactcattca gtgtgccaag
     3721 gacatcgcca aggcctcaga tgaggtgact cggttggcca aggaggttgc caagcagtgc
     3781 acagataaac ggattagaac caacctctta caggtatgtg agcgaatccc aaccataagc
     3841 acccagctca aaatcctgtc cacagtgaag gccaccatgc tgggccggac caacatcagt
     3901 gatgaggagt ctgagcaggc cacagagatg ctggttcaca atgcccagaa cctcatgcag
     3961 tctgtgaagg agactgtgcg ggaagctgaa gctgcttcaa tcaaaattcg aacagatgct
     4021 ggatttacac tgcgctgggt tagaaagact ccctggtacc agtgagcggc cgcatcgtga
     4081 ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4141 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4201 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     4261 tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     4321 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4381 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4441 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4501 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4561 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4621 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4681 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     4741 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4801 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4861 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4921 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4981 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     5041 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5101 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5161 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5221 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5281 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5341 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5401 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5461 tgagttttcg ttccactgag cgtcagaccc cg
//
