LOCUS       sequence-417684-        7119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a tandem dimer of mVenus in zebrafish pancreatic beta 
            cells using a zebrafish insulin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7119)
  AUTHORS   Bui CV, Boswell CW, Ciruna B, Rocheleau JV
  TITLE     Apollo-NADP(+) reveals in vivo adaptation of NADPH/NADP(+) 
            metabolism in electrically activated pancreatic beta cells.
  JOURNAL   Sci Adv. 2023 Oct 6;9(40):eadi8317. doi: 10.1126/sciadv.adi8317. 
            Epub 2023 Oct 4.
  PUBMED    37792934
REFERENCE   2  (bases 1 to 7119)
  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: "Sci Adv."; 
            date: "2023-10-6"; volume: "9(40):eadi8317. doi"; pages: " 
            10.1126/sciadv.adi8317. Epub 2023 Oct 4"
COMMENT     Sequence Label: pDestTol2pA ins:TDimer mVenus
FEATURES             Location/Qualifiers
     source          1..7119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(124..712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(204..223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(883..1674,1675..1743))
                     /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     1506..1525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1744..1848)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2011..2032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(2381..2392,2393..2555)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     2928..2944
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2964..2984
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    4141..4165
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4183..4192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4189..4191,4192..4194,4195..4905)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      4921..4930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4927..4929,4930..4932,4933..5643)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(5660..5684)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5686..5820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(5883..5903)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(5911..5927)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5943..6077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(6516..6703,6704..6715)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6749..6765)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6749..6765)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6802..6822)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6802..6820)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6841..6857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6854..6876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6865..6881
                     /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(6889..6895,6896..6913,6914..6919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6934..6955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7072..7089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
       61 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      121 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      181 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      241 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      301 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      361 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      421 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      481 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      541 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      601 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      661 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      721 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aaggcatttt
      781 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      841 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      901 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      961 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1021 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1081 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1141 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1201 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1261 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1321 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1381 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1441 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1501 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1561 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1621 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1681 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1741 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1801 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1861 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     1921 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     1981 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2041 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccacgctgg
     2101 ctaagaactc atcagcctcc ccggtccatc tacccacgta ccaatgcacc aattggccac
     2161 aatgacggct actacatggt gccattcctt cctctttata ggaatggaga ctacctcctg
     2221 tccaacaatg ctcttggata cgagtacgcc tacctgttgg acccaggtca ttgcacaaca
     2281 ccagaaatgc cctctgatct gcaaaagacg tgaatatctg ttcagacacc catatccact
     2341 ctgttccaca caggtcagag gtttgtccag gagttcttga cagaggtgta aaaagtactc
     2401 aaaaatttta ctcaagtgaa agtacaagta cttagggaaa attttactca attaaaagta
     2461 aaagtatctg gctagaatct tacttgagta aaagtaaaaa agtactccat taaaattgta
     2521 cttgagtatt aaggaagtaa aagtaaaagc aagaaagaaa actagagatt cttgtttaag
     2581 cttttaatct caaaaaacat taaatgaaat gcatacaagg ttttatcctg ctttagaact
     2641 gtttgtattt aattatcaaa ctataagaca gacaatctaa tgccagtaca cgctactcaa
     2701 agttgtaaaa cctcagattt aacttcagta gaagctgatt ctcaaaattg ttagtgtcaa
     2761 gcctagctct tttggggctg aaaagcaatc ctgcagtgct gaaaagcctc tcacaggcag
     2821 ccgatgcggg aagaggtgta ttagtcttga tagagaggct gcaaatagca ggaaacgtga
     2881 gcagagactc cctggtgtct gaaacacagg ccagatgggc cctcgagcag gaaacagcta
     2941 tgaccatgat tacgccaagc tatcaacttt gtatagaaaa gttgtagagc tcacgcgtaa
     3001 gctttgtttg taccataata agctcattat ataatattta aaatgatact gcatcattaa
     3061 ttgatatgtt ttctagttat ttttaaatat tagcaactaa attaggaaat tatccatagt
     3121 aaatttaatg taatattatt tgttatattt aacttcagcc cacagtctag tttagttgtt
     3181 ggtcccttat gagaaaaaat ttgggcacct ctgctctagt gcttgtaaac atttttgagt
     3241 atctatagtt gaacatgaaa gcattttttt tttttttttt ttaagaaaat gaagaaagct
     3301 aaaaacctgt atccactgac ttccattaaa aaaagcaaat actagggatg tctatggtgt
     3361 cttcttttgt gttcaacaaa taagcagaaa gtaactgctg acagaacatt acattttgtg
     3421 tgaatcccct tatattgtgt taaaaactgt agagaaatca tctaataata agaaattatc
     3481 aagaaataaa cagaaaatga tctaatagta tactataaaa tctttcaaaa tccaggccac
     3541 taccatatga ctaaaaccac acatattatg ctttaaaaaa agtcacaatg aagacgtcac
     3601 aaacgttcct gaagactaat gactcgtcta accccatctt accttctgct gtttcactca
     3661 aagaaaagcc accagctgtc tctgtgatct gctcattctg tgttaacttg tgttctcaac
     3721 aattaacaat taaccaaaag cctctggagc tgcgtgagca tcacgatgtg tttgtcttct
     3781 gtgagcgtgg cagtgtccac gaagtcaatg tgaattggtt ggccaaacac agctaatgtg
     3841 gcatgtgtga ttggacagcc acttaaaagt gttaaagtgt gtttacgcac aggattggtt
     3901 ttccgctgaa tcaggcaaaa ccagcagaac cgatctaaac tcgagtataa aagtgagtag
     3961 gagatgctat tcgtcccaaa acatctccac caccatctcc accattcctc gcctctgctt
     4021 cgagaacagg tgagtgtcga gcgggatggt aaatctacag agaatgcgga gtgtagcttg
     4081 tgtacatgtt tttgattaac agagattgta tgtgtgtgtt tgtgtcagtg tgacggatcc
     4141 gcaagtttgt acaaaaaagc aggcttcagt gtggtggaat tcgccaccat ggtgagcaag
     4201 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     4261 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     4321 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     4381 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     4441 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     4501 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     4561 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     4621 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     4681 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     4741 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     4801 cagtccaaac tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     4861 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagatgga tccaccggtc
     4921 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4981 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     5041 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     5101 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
     5161 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     5221 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     5281 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     5341 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     5401 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     5461 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     5521 aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac
     5581 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     5641 aagctcgagt ctagagggca cccagctttc ttgtacaaag tgggggatcc agacatgata
     5701 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     5761 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     5821 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     5881 tccaacttta ttatacatag ttgataattc actggccgtc gttttacggt accatcgatg
     5941 atgatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     6001 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     6061 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     6121 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     6181 atcctctaga tcagatctgc gaagatacgg ccacgggtgc tcttgatcct gtggctgatt
     6241 ttggactgtg ctgctcgcag ctgctgatga atcacatact tcctccattt tcttccactg
     6301 attgactgtt ataatttccc taatttccag gtcaaggtgc tgtgcattgt ggtaatagat
     6361 gtgacatgac gtcacttcca aaggaccaat gaacatgtct gaccaatttc atataatgtg
     6421 aaaacgattt tcataggcag aataaataac atttaaatta aactgggcat cagcgcaatt
     6481 caattggttt ggtaatagca agggaaaata gaatgaagtg atctccaaaa aataagtact
     6541 ttttgactgt aaataaaatt gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa
     6601 ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa
     6661 aataatactt aagtacagta atcaagtaaa attactcaag tactttacac ctctggttct
     6721 tgacccccta ccttcagcaa gcccagcaga tccactagtt ctagagcggc cgccaccgcg
     6781 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     6841 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6901 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6961 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7021 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7081 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggt
//
