LOCUS       sequence-363552-        8031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP tagged form of zebrafish vinculin b. 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 8031)
  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 8031)
  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-vclb
FEATURES             Location/Qualifiers
     source          1..8031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..45
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     27..44
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(83..85,86..88,89..799)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(83..85,86..88,89..799)
                     /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(128..149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(389..408)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     736..757
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4075..4209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4099..4118)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4153..4172
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4322..4342)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4322..4340)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4361..4377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4361..4377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4374..4396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4385..4401
                     /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(4409..4415,4416..4433,4434..4439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4454..4475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4592..4609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4763..5351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4843..4862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5522..6313,6314..6382))
                     /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     6145..6164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6383..6487)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6513..6968)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6650..6671)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6862..6881
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7039..8023
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt
       61 tctttttgca ggatccgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      121 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      181 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      241 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      301 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      361 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      421 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      481 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      541 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
      601 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      661 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
      721 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
      781 catggacgag ctgtacaagt actcagatct cgagctcaag cttatgccgg ttttccacac
      841 gaagacgatc gagagtatcc tggagccggt ggcccagcag atctcccatc tggtgatcat
      901 gcacgaggag ggggaggtgg acgggaaagc catcccggac ctcagcgctc ctgtgttggc
      961 cgtgcaggcg gctgtcagca acctggtccg ggtgggcaaa gagacggttc agaccactga
     1021 ggatgccatc atgaggagag acatgccacc cgccttcata aaggtggaga acgcgtgcac
     1081 taaactggtt caggcggctc agatgctgaa ggccgacccg tattcagtgc cggctagaga
     1141 ttatctgatc gacggctccc gtgggatcct gtccggcacg tctgacctcc tgctgacctt
     1201 cgacgaggcc gaggtccgta aaatcatccg cgtgtgtaaa ggcattctgg agtatctgac
     1261 cgtagcagag gtggtggagt ccatggagga tctcatcact tacaccaaaa acctgggacc
     1321 cggcatgacg aagatggcca agatgattga tgagcgccag caggaattaa cccatcagga
     1381 gcacagagtc atgctggtca actccatgaa cactgtgaag gagctcctgc ccgtcctcat
     1441 ctcaggcatc aagatcttcg taaccaccag gacgtctcag ggaaagggtg tggaggaggc
     1501 gctgaagaac aggaacttca cagtggagaa gatgaacaca gaaatcaacg agatcatccg
     1561 cgttctgcag ctcacgtcct gggatgagga tgcctgggcc aataaggaca cagaggccat
     1621 gaagcgcgcg ctggctctca ttgactccaa aatggcccaa gccaaaaact ggctccgaga
     1681 ccctcaaggc caaccaggag gtccaggtga acaggccata cggcagatct tagatgaagc
     1741 tgagaaagtt ggtgaactct gtgctgggaa agagagacga gacattgtgg gaacagccaa
     1801 aacactcgga caacttacag aacaggtgtc tgatctaagg gccaggggtc agggtgcgaa
     1861 tccagttgcc atgcaaaaag cccagcaggt ctctcagggt ctggatgtcc tgacagggaa
     1921 agtggaaaac gcagctcgaa aactggaggc aatgaccggc tccaaacagg ccatcgctaa
     1981 gaggatcgat gcggctcaga gctggcttgc agatcctcat agtggcccag aaggagagga
     2041 gaatatcagg gctcttctag gagaagccag gaagattgca gatctgtgtg aagatcccaa
     2101 agagcgggag gatatcctgc gcagcatgag tgagatcgca gcactcagcg ggaaactagc
     2161 agagctcaag agagcaggta aaggtgatac tccagaggct cgagctctgg ctaagcagat
     2221 cgccacggct ctgcagaacc tgcagtctaa gaccagtaaa gcggtggcaa acactcgacc
     2281 ggccaaagca gccgttcatc tggccggaaa gatggagcag gccggccgct ggattgacaa
     2341 ccccacgctg gacgacagtg gcgtcggtca ggctgccatc cgcggtctgc tggcagaggg
     2401 tcgtcgtctg gccaatgctc tgccggcggc gcagcgtcag gagctgctgg gtaaatgtga
     2461 gcaggttgaa cacctgatgg cacagctggc agagctggca gctcgaggag aaggagacag
     2521 tccacaggcc cgtgccatcg ctcagcagct gcaacacaca ctcaaagagc tggagggtaa
     2581 gatgcaagac gccatgacac aggaagtgtc cgacatcttc agcgacacca ccactccaat
     2641 caaactactg gctgttgcgg cgacggctcc ttcagactct cccaacagag aggaggtgtt
     2701 taaggaacgg gcctcaaact ttgagaatca cgctagtcgt ctgggagcca cggcggagaa
     2761 agcagcagca gtcgggaccg caaacaagag cacggtggag ggcatacagg cggcggtcaa
     2821 atcagccaga gacctgacgc cgcaggtcgt ctctgctgct cgcatcctgc tgaagaatcc
     2881 tggaaatcag gcagcgtttg agcacttcga gaccatgaag aaccagtgga ttgacaacgt
     2941 ggagaaaatg acaggtctgg tggatgaagc catagacacc aggtctctgc tcgccgcatc
     3001 ggaagacgcc attaagaaag atctggataa atgtcaggtg gccatggcca atcaccagcc
     3061 tcagatgctt gtcgccgggg caaccagcat cgcccgacgg gccaatcgga tcctgctggt
     3121 ggcgaagcga gagattgaaa actcagagga cccgaagttc agagaaacgg tgaaagcggc
     3181 ttcagacgag ctcagcagaa ccatctctcc aatggtcatg gatgccaaag ctgtggccgc
     3241 aaacataaag gaccaaggcc ttcagagggg cttcctggac tccggtttta agattctggg
     3301 tgcggtggct aacgtcaggg acgctttcca gccgcaggag cccgaattcc ctcctccacc
     3361 tccagacctg gagagcctgc agataagcga cacggctgct cctcccaaac caccgctgcc
     3421 ggagggtgaa gttcctccac ccagacctcc accacccgag gagaaggacg aggagttccc
     3481 ggagcagcag gccggagaga tggtcagcga gcccatgatg gtggccgcac gacagctgca
     3541 cgacgaggcc cgcaagtggt ccagcaaggg taatgatatc attggtgcgg cgaagcgcat
     3601 ggctctgctg atggctgaga tgtctcgtct ggtccggggc agtggcggaa ataagagagc
     3661 cctcatccag tgcgctaaag acattgctaa agcctctgat gaggtcacgc gcctcgctaa
     3721 agaggttgct aaacagtgca cagacaaacg catacggacc aacctgctgc aggtgtgtga
     3781 gaggatcccc accatcagta ctcagctgaa gattctgtcc acggtgaagg ctacaatgtt
     3841 gggccgcacc aacattagcg aggaggaatc agagcaggcc actgagatgc tggtccacaa
     3901 cgctcagaac ctcatgcagt ccgtgaaaga gactgtccga gaagctgaag ccgcatccat
     3961 caagatccgc actgacgccg gcttcaccct gcactgggtc cgcaagacac cctggtacca
     4021 gtgagaattc aaggcctctc gagcctctag aactatagtg agtcgtatta cgtagatcca
     4081 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4141 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4201 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     4261 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     4321 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4381 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     4441 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4501 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4561 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4621 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4681 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4741 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4801 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4861 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4921 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4981 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5041 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5101 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5161 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5221 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5281 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5341 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5401 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5461 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5521 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5581 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5641 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5701 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5761 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5821 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5881 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5941 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6001 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6061 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6121 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6181 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6241 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6301 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6361 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6481 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6541 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6601 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6661 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6721 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6781 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6841 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6901 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6961 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7021 ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa
     7081 ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa
     7141 ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa
     7201 ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca
     7261 agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca
     7321 ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc
     7381 cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     7441 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     7501 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     7561 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     7621 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     7681 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccgttctca
     7741 cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt
     7801 aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc
     7861 ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg
     7921 tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg
     7981 tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aaccgccatt c
//
