LOCUS       sequence-410265-        8127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mVenus-tagged Apollo-NADP+ 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 8127)
  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 8127)
  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:Apollo-NADP+ mVenus
FEATURES             Location/Qualifiers
     source          1..8127
                     /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"
     regulatory      4159..4168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5719..5728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5725..5727,5728..5730,5731..6444)
                     /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"
     primer_bind     complement(5770..5791)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6031..6050)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6378..6399
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6473..6497)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6568..6691)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    6694..6828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6891..6911)
                     /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(6919..6935)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6951..7085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(7524..7711,7712..7723)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(7757..7773)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7757..7773)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7810..7830)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7810..7828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7849..7865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7862..7884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7873..7889
                     /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(7897..7903,7904..7921,7922..7927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7942..7963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8080..8097)
                     /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 gaaggacagt atttggtatc tgcgctctgc tgaagccagt
      601 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      661 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      721 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      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 tccaacaagg 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 cccgggctgc aggaattcgc caccatggca gagcaggtgg ccctgagccg gacccaggtg
     4201 tgcgggatcc tgcgggaaga gcttttccag ggcgatgcct tccatcagtc ggatacacac
     4261 atattcatca tcatgggtgc agcgggtgac ctggccaaga agaagatcta ccccaccatc
     4321 tggtggctgt tccgggatgg ccttctgccc gaaaacacct tcatcgtggg ctatgctcag
     4381 tcccgcctca cagtggctga catccgcaaa cagagtgagc ccttcttcaa ggccacccca
     4441 gaggagaagc tcaagctgga ggacttcttt gcccgcaact cctatgtggc tggccagtac
     4501 gatgatgcag cctcctacca gcgcctcaac agccacatga atgccctcca cctggggtca
     4561 caggccaacc gcctcttcta cctggccttg cccccgaccg tctacgaggc cgtcaccaag
     4621 aacattcacg agtcctgcat gagccagata ggctggaacc gcatcatcgt ggagaagccc
     4681 ttcgggaggg acctgcagag ctctgaccgg ctgtccaacc acatctcctc cctgttccgt
     4741 gaggaccaga tctaccgcat cgacaactac ctgggcacgg agatggtgca gaacctcatg
     4801 gtgctgagat ttgccaacag gatcttcggc cccatctgga accgggacaa catcgcctgc
     4861 gttatcctca ccttcaagga gccctttggc actgagggtc gcgggggcta tttcgatgaa
     4921 tttgggatca tccgggacgt gatgcagaac cacctactgc agatgctgtg tctggtggcc
     4981 atggagaagc ccgcctccac caactcagat gacgtccgtg atgagaaggt caaggtgttg
     5041 aaatgcatct cagaggtgca ggccaacaat gtggtcctgg gccagtacgt ggggaacccc
     5101 gatggagagg gcgaggccac caaagggtac ctggacgacc ccacggtgcc ccgcgggtcc
     5161 accaccgcca cttttgcagc cgtcgtcctc tatgtggaga atgagaggtg ggatggggtg
     5221 cccttcatcc tgcgctgcgg caaggccctg aacgagcgca aggccgaggt gaggctgcag
     5281 ttccatgatg tggccggcga catcttccac cagcagtgca agcgcaacga gctggtgatc
     5341 cgcgtgcagc ccaacgaggc cgtgtacacc aagatgatga ccaagaagcc gggcatgttc
     5401 ttcaaccccg aggagtcgga gctggacctg acctacggca acagatacaa gaacgtgaag
     5461 ctccctgacg cctatgagcg cctcatcctg gacgtcttct gcgggagcca gatgcacttc
     5521 gtgcgcagcg acgagctccg tgaggcctgg cgtattttca ccccactgct gcaccagatt
     5581 gagctggaga agcccaagcc catcccctat atttatggca gccgaggccc cacggaggca
     5641 gacgagctga tgaagagagt gggtttccag tatgagggca cctacaagtg ggtgaacccc
     5701 cacaagctca gaccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     5761 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     5821 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     5881 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
     5941 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     6001 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     6061 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     6121 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     6181 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     6241 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     6301 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaaactgag caaagacccc
     6361 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     6421 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc acacccagct
     6481 ttcttgtaca aagtggtctt aagctcgggc cctgcagctc tagagctcga attctacagg
     6541 tcactaatac catctaagta gttggttcat agtgactgca tatgttgtgt tttacagtat
     6601 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
     6661 gtttctcgtt caactttctt gtacaaagtg ggggatccag acatgataag atacattgat
     6721 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     6781 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     6841 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc caactttatt
     6901 atacatagtt gataattcac tggccgtcgt tttacggtac catcgatgat gatccagaca
     6961 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     7021 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     7081 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     7141 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat cctctagatc
     7201 agatctgcga agatacggcc acgggtgctc ttgatcctgt ggctgatttt ggactgtgct
     7261 gctcgcagct gctgatgaat cacatacttc ctccattttc ttccactgat tgactgttat
     7321 aatttcccta atttccaggt caaggtgctg tgcattgtgg taatagatgt gacatgacgt
     7381 cacttccaaa ggaccaatga acatgtctga ccaatttcat ataatgtgaa aacgattttc
     7441 ataggcagaa taaataacat ttaaattaaa ctgggcatca gcgcaattca attggtttgg
     7501 taatagcaag ggaaaataga atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa
     7561 ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt
     7621 aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa
     7681 gtacagtaat caagtaaaat tactcaagta ctttacacct ctggttcttg accccctacc
     7741 ttcagcaagc ccagcagatc cactagttct agagcggccg ccaccgcggt ggagctccag
     7801 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     7861 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7921 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7981 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     8041 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     8101 ctcggtcgtt cggctgcggc gagcggt
//
