LOCUS       sequence-363548-        8032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP tagged form of zebrafish vinculin a. For in vitro transcription 
            (SP6) or expression through CMV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8032)
  AUTHORS   Boutillon A, Escot S, Elouin A, Jahn D, Gonzalez-Tirado S, Starruss 
            J, Brusch L, David NB
  TITLE     Guidance by followers ensures long-range coordination of cell 
            migration through alpha-catenin mechanoperception.
  JOURNAL   Dev Cell. 2022 May 17. pii: S1534-5807(22)00329-X. doi: 
            10.1016/j.devcel.2022.05.001.
  PUBMED    35613615
REFERENCE   2  (bases 1 to 8032)
  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"; journalName: "Dev Cell. 
            2022 May 17. pii: S1534-5807(22)00329-X. doi: 
            10.1016/j.devcel.2022.05.001."
COMMENT     Sequence Label: pCS2+ Zf GFP-vcla
FEATURES             Location/Qualifiers
     source          1..8032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..65
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     47..64
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      96..105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(102..104,105..107,108..818)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(102..104,105..107,108..818)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(408..427)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     755..776
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4096..4230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4120..4139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4174..4193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4343..4363)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4343..4361)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4382..4398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4382..4398)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4395..4417)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4406..4422
                     /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)."
     promoter        complement(join(4430..4436,4437..4454,4455..4460))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4475..4496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4613..4630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4784..5372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4864..4883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5543..6334,6335..6403))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6166..6185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6404..6508)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6534..6989)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6671..6692)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6883..6902
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(7060..8032,1..12)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgtttaggga accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact
       61 atagaataca agctacttgt tctttttgca ggatcgccac catggtgagc aagggcgagg
      121 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      181 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      241 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      301 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      361 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      421 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      481 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      541 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
      601 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
      661 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
      721 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
      781 ccgccgggat cactctcggc atggacgagc tgtacaagta ctcagatctc gagctcaagc
      841 ttatgccagt tttccatacc aagacgatag agagtatcct ggagccggtg gctcagcaga
      901 tttcacactt ggttatcatg cacgaggagg gagaagttga tgggaaagcg atcccagatc
      961 tcaccagccc agtggcggca gtgcaagcgg ctgtcagcaa ccttgtgcgg gtggggaagg
     1021 agacagtgca gaccacagaa gacaagatta tgaagagaga catgccatca gccttcatta
     1081 aggtggagaa cgcctgtgca aagctggtgg aagcagctca gatgctgaga acagacccgt
     1141 actctgtccc ggcccgtgat tacctcatcg acggctctcg tggaatcctc tcaggcacct
     1201 cagacctgct gctgaccttc gatgaagccg aggtgcgtaa aattatccgc gtgtgtaagg
     1261 gcatcctgga atacctgact gttgcagagg ttgtggaaac catggaggac ttgatcactt
     1321 acactaaaaa cctgggacca ggcatgacga agatggcaaa gatgattgac gagcggcagc
     1381 aggaactcac gcaccaggag caccgcgtaa tgttggtcac ctccatgaac acagtaaagg
     1441 agctgctgcc cgtgcttata tcagccatta aaatctttgt gacaaccaag tgcactaaga
     1501 gtcacggtgt ggaggaggcc ttgaagaaca gaaactacac gtttgataag atgacagctg
     1561 aaatcaacga gataatcaga gtgctgcagc tcacttcatg ggacgaggac gcctgggcaa
     1621 ataaaaagga cactgaggcc atgaagagag cgctggcgct gatcgagtct aagatgggtc
     1681 aggctaaagg ctggctgagg gaccccaatg cactgccagg agatccagga gagcatgcgc
     1741 tgcgtcagat cctggatgaa gcaggaaaag tgggtgagct ttgtgcaggg aaagagagac
     1801 gagagatcct gggaacagcc aaaaccctgg gccagatgac ggatcaggtg tcagatgtgc
     1861 gtgccagagg tcagggtgcc acacctatgg gtatgcagaa agctcagcag gttgctcaag
     1921 gtttggatat tctggtggga aaagtggaga acgcagcccg taagctggag gccctaacca
     1981 atgccaaaca ggccatcgcc aagagaatcg acaatgccca gagctggctg gctgatccta
     2041 acggcggtcc agagggtgaa gaaaacatcc gcgctcttct ggcggaagcc aaacgcatcg
     2101 ctgacttgtg tgaagaccct aaagaacgag atgacattct tcgttccatc ggcgagatcg
     2161 caggcctcac cgccaggctg gtcgagctcc gtagaatagg taaaggcgat actccagagg
     2221 cgagagctct tgccaaacag attggcacag ctttgcagaa cttacaggca aaaacgaacc
     2281 gtgctgtggc caacatgagg cctgccaaag ctgctgttac cctggagagc aaaatggagc
     2341 aggccttgcg ctggatcaac aaccctggag tggacgatca tggagtaggc caggctgcta
     2401 tacgagggct gatagcagag ggtcgacggc tcgcgagctc tctgccaggg ccgtacagac
     2461 aggagctgct ggccaaatgt gagcaggtgg agcagctgat gatgcagctg gcggatctgg
     2521 cagcgcgggg agaaggagag agtcctcagg cccgcgcagt ggctgctcat cttctggagg
     2581 caattaaaga tctgaaggca aagatgcagg aagcaatgac ccaggaagtg tctgatgttt
     2641 tcagcgatac aacaacacca attaaactcc tggctgtggc agcaacagca cctctcgagg
     2701 ccccgaacag agaggaggtg tttgaagaaa gggcttctaa ctttgagaat catgctagca
     2761 ggctgggtgc tactgcggag aaggcggctg cggtaggaac agccaataaa agcacagttg
     2821 aaggcatcca agcggctgtg aaatcagcta gagatctaac acctcaggtc acttccgctg
     2881 cacgtatttt gctgaagaac cctggcaacc aggccgccta cgagcacttt gataccatga
     2941 agaaccagtg gattgacaac attgaaaaaa tgaccagtct tgtggatgaa gccattgaca
     3001 ctaaatccct cttggatgct tcagaggaag ctattaagaa agacattgat aaatgccgtg
     3061 tagcaatggc taacgttcag ccccagatgt tggttgcagg ggctacaagc attgcccggc
     3121 gggcaaacag agtacttctg gtggccaaga gggaagtgga gaattcagag gaccctaaat
     3181 tcagagaact ggttaaagca gcttcagatg aactcggtag aaccatctca cccatggtta
     3241 tggctgctaa aggagtggca ggaaacatcc aggatccagg tttgcaaaag ggcttcctgg
     3301 actctggcta caggatcttg gctgctgttg ggaaagtcag agaagccttt cagcctcagg
     3361 agctggactt tcctccgcca cctccagacc tggatcagct gcatgtcaat gatgaccagg
     3421 ctccacccaa accacctctt cctgagggag aggttcctcc gccgagacct ccacctccag
     3481 aagagaaaga tgaggagttc cctgagcaga aggcaggaga gatggtgagc gagcccatga
     3541 tggtggctgc caggtcactg cacgacgagg cccgcaagtg gtccagcaag ggtaatgaca
     3601 tcattggagc cgccaagcgt atggccctgc tgatggcaga gatgtctcgt ctggtgcgtg
     3661 gtggaagtgg gaataaacgt gcccttattc agtgcgccaa ggacatcgcc aaggcctctg
     3721 atgaggtcac gcggctggct aaagaagtag ccaaacaatg tactgacaaa aggattcgca
     3781 caaacctgct gcaggtttgt gagcgcatcc caaccatcag cactcagttg aagatccttt
     3841 ccacagtgaa ggccactatg ctgggacgta caaatatcag tgaggaagaa tccgaacagg
     3901 ccactgagat gcttgtgcac aacgcccaga acctcatgca gtctgtgaag gagacggtga
     3961 gagaagcaga agccgcttcc atcaagatcc gcacagacgc aggcttcaca cttcgctggg
     4021 tccgaaaaac cccctggtac cagtaaaatt caaggcctct cgagcctcta gaactatagt
     4081 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     4141 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4201 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4261 caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg
     4321 gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     4381 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4441 ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4501 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4561 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4621 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4681 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4741 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4801 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4861 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4921 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4981 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5041 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5101 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5161 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5221 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5281 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5341 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5401 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5461 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5521 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5581 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5641 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5701 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5761 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5821 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5881 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5941 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6001 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6061 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6121 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6181 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6241 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6301 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6361 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6421 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6481 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     6541 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6601 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6661 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6721 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6781 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6841 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6901 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6961 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     7021 ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg
     7081 gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg
     7141 gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg
     7201 gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg
     7261 ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca
     7321 tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat
     7381 ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg
     7441 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
     7501 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
     7561 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
     7621 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
     7681 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
     7741 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
     7801 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
     7861 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
     7921 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
     7981 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ct
//
