LOCUS       sequence-414384-        8512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses membrane-labelled mCherry in the zebrafish heart.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8512)
  AUTHORS   Jia BZ, Qi Y, Wong-Campos JD, Megason SG, Cohen AE
  TITLE     A bioelectrical phase transition patterns the first vertebrate 
            heartbeats.
  JOURNAL   Nature. 2023 Oct;622(7981):149-155. doi: 10.1038/s41586-023-06561-z.
            Epub 2023 Sep 27.
  PUBMED    37758945
REFERENCE   2  (bases 1 to 8512)
  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: 
            "10.1038/s41586-023-06561-z"; journalName: "Nature"; date: 
            "2023-10"; volume: "622"; issue: "7981"; pages: "149-155"
COMMENT     Sequence Label: pBJ077_zACNE20_E1b_2xLyn_mCherry
FEATURES             Location/Qualifiers
     source          1..8512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(300..462,463..474)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2325..2344)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2326..2344)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2354..2371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2354..2370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2512..2967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2599..2618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2809..2830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2993..3097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3098..3166,3167..3958)
                     /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     complement(3316..3335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4129..4717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4618..4637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4871..4888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5005..5026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5041..5046,5047..5064,5065..5071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5079..5095
                     /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     5084..5106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5103..5119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5103..5119
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5138..5158
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5140..5158
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     5195..5211
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5195..5211
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(5245..5256,5257..5444)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    5881..6015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5918..5937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5972..5991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6040..6750)
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     6587..6605
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6825..6834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      6879..6888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          6962..7534
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    7642..7666
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(8409..8433)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 tctatcaaga ctaatacacc tcttcccgca tcggctgcct gtgagaggct tttcagcact
       61 gcaggattgc ttttcagccc caaaagagct aggcttgaca ctaacaattt tgagaatcag
      121 cttctactga agttaaatct gaggttttac aactttgagt agcgtgtact ggcattagat
      181 tgtctgtctt atagtttgat aattaaatac aaacagttct aaagcaggat aaaaccttgt
      241 atgcatttca tttaatgttt tttgagatta aaagcttaaa caagaatctc tagttttctt
      301 tcttgctttt acttttactt ccttaatact caagtacaat tttaatggag tactttttta
      361 cttttactca agtaagattc tagccagata cttttacttt taattgagta aaattttccc
      421 taagtacttg tactttcact tgagtaaaat ttttgagtac tttttacacc tctgtcaaga
      481 actcctggac aaacctctga cctgtgtgga acagagtgga tatgggtgtc tgaacagata
      541 ttcacgtctt ttgcagatca gagggcattt ctggtgttgt gcaatgacct gggtccaaca
      601 ggtaggcgta ctcgtatcca agagcattgt tggacaggag gtagtctcca ttcctataaa
      661 gaggaaggaa tggcaccatg tagtagccgt cattgtggcc aattggtgca ttggtacgtg
      721 ggtagatgga ccggggaggc tgatgagttc ttagccagcg ctcaaagatg ctagaaatga
      781 tgtaagaaac aaatatgcaa gatgaaagtt aagataagat aaaaaattaa acaaataaat
      841 gcaattatgt tttgtgtttc tattccacca ataataacct cacgcagtaa agccaccaga
      901 ttattttttg ttctttgctc tgcagggcaa ccaaactcta acaattgatc tcaattgatc
      961 tttcgctgct catccaagta cattcacaaa cttcactaat tgtagcagct ataaacaaca
     1021 gattgtatga actttgtcaa aaactgtgca gtctaatgtc atgacgtgtt tgttacctgt
     1081 caatgaaagc atggtgaagg aggaagatgg ggtcgttggc tgagccctgg actgaggaca
     1141 ttgagccgtt catgaagaca tgcagcgcat tgtgcatggt gctctggcct tgcacagcca
     1201 tgcctgtttc tgggctggca aaacctacaa agaaaagaag caggtgaaga agttacagtt
     1261 tcaatcaaga taacagcaaa acaaggctaa ttaaaagaca gcaaaaaaag gagatgtaaa
     1321 atatagctat tgattttctt tacttttctt tacgtaggca aatgtgactt aagtgtgttg
     1381 ttgtatcccc cccccccccc agttttttat gtatgcattt aatcatttct gtaatgaatc
     1441 tgtggaaatt atttcttatt ttgactacca atcaatcaat caatcaatca atcaatcaat
     1501 caatcaatca atcaatcaat caatcaatca atcaatcaat caatctacca accaaccaac
     1561 caaccaacga ctgtatatga gaactgtaat gagtgacctc ctccttctag tgatccaaac
     1621 aggaagaagc tccaaatggc atgaatgctc cagtggtggc attcatatta aaatctttat
     1681 ttaacatgta cttgttaatc ataatttctg taataacttt tcttttctgt agcccaagtt
     1741 gttgatatta taaactgacc aaccagatgc cttaataaaa gtatgtgctc tcacatccaa
     1801 cgttcaaaac gtttgacaga ttttgtgcag cccgctttca agtggtaggc tgtggacttc
     1861 caacaagctt acttctgatt ggttagaggg gttgccatag gaacggtgac tcagactcga
     1921 cgcatctcgg ctctttcttg cttattggca acgtctagct ccaacacggc gatgtccata
     1981 cagtgaaaaa tggcaactga attaacttca ttttgttgga gccagaagta agtcattttg
     2041 aatgggtgac gtaacactca ctcggtccag ttctcaaata cagtcaacaa gtactattac
     2101 ataataatag tgtgtactac tattacatat tattgtaata atgtctgtta aacacagtca
     2161 taattgaaaa aaaacaacta aaacatttat tcaagggtaa agacaaatgc agacaaatac
     2221 agaaaagatt taaaatagca gtagaaaatg gcaaacgggc ttattcatcc aagttttcag
     2281 cacttgtaca ggtgggtaat tcctactatt tggtacccaa ttcgccctat agtgagtcgt
     2341 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2401 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2461 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2521 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2581 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2641 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2701 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2761 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2821 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2881 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2941 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3001 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3181 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3241 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3301 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3361 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3421 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3481 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3541 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3601 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3661 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3721 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3781 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3841 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3901 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3961 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4021 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4081 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4261 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4501 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4741 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4801 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4861 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4921 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4981 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5041 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5101 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     5161 acaaaagctg gagctccacc gcggtggcgg ccgctctaga actagtggat ctgctgggct
     5221 tgctgaaggt agggggtcaa gaaccagagg tgtaaagtac ttgagtaatt ttacttgatt
     5281 actgtactta agtattattt ttggggattt ttactttact tgagtacaat taaaaatcaa
     5341 tacttttact tttacttaat tacatttttt tagaaaaaaa agtacttttt actccttaca
     5401 attttattta cagtcaaaaa gtacttattt tttggagatc acttcattct attttccctt
     5461 gctattacca aaccaattga attgcgctga tgcccagttt aatttaaatg ttatttattc
     5521 tgcctatgaa aatcgttttc acattatatg aaattggtca gacatgttca ttggtccttt
     5581 ggaagtgacg tcatgtcaca tctattacca caatgcacag caccttgacc tggaaattag
     5641 ggaaattata acagtcaatc agtggaagaa aatggaggaa gtatgtgatt catcagcagc
     5701 tgcgagcagc acagtccaaa atcagccaca ggatcaagag cacccgtggc cgtatcttcg
     5761 gatctgatct agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     5821 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     5881 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5941 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6001 tatcatgtct ggatcatcat cgatgaatta attcgagctt cacttgtaca gctcgtccat
     6061 gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc
     6121 gttgtgggaa aggatgtcca acttgatgtc gacgttgtag gcgccgggca gctgcacggg
     6181 cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt
     6241 cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcagtgga
     6301 ggcctcccag cccatggtac gacactgcat tacggggccg tcggagggga agttggtgcc
     6361 gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt
     6421 caccacgccg ccgtcctcga agttcatcac gcgctcccaa ttgaagccct cggggaagga
     6481 cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat
     6541 gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag
     6601 cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc
     6661 gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc
     6721 catgttatcc tcctcgccct tgctcaccat tgctgcacct gcacctccag ctggcgctcc
     6781 ctcgtcgtcg ttcaggttgt ccttgcgctt gctcttgatg cagcccatgg cggcctcgtc
     6841 gtcgttcagg ttgtccttgc gcttgctctt gatgcagccc atggtggcga ccggtggatc
     6901 cgtcgaggaa ttctttgcca aaatgatgag acagcacaac aaccagcacg ttgcccagga
     6961 gctgtaggaa agagaagaag gcatgaacat ggttagcaga ggggcccggt ttggactcag
     7021 agtattttat cctcatctca aacagtgtat atcattgtaa ccataaagag aaaggcagga
     7081 tgatgaccag ggtgtagttg tttctaccaa taagaatatt tccacgccag ccagaattta
     7141 tatgcagaaa tattctacct tatcatttaa ttataacaat tgttctctaa aactgtgctg
     7201 aagtacaata taatataccc tgattgcctt gaaaaaaaag tgattagaga aagtacttac
     7261 aatctgacaa ataaacaaaa gtgaatttaa aaattcgtta caaatgcaag ctaaagttta
     7321 acgaaaaagt tacagaaaat gaaaagaaaa taagaggaga caatggttgt caacagagta
     7381 gaaagtgaaa gaaacaaaat tatcatgagg gtccatggtg atacaaggga catcttccca
     7441 ttctaaacaa caccctgaaa actttgcccc ctccatataa catgaatttt acaatagcga
     7501 aaaagaaaga acaatcaagg gtccccaaac tcaccctgaa gttctcagga tcggtcgaaa
     7561 ttcgtgtgga ggagctcaaa gtgaggctga gacgcgatgg gatccattat ataccctcta
     7621 gagtcgagag atcttgatat caccactttg tacaagaaag ctgggtgtgg tagaaagtaa
     7681 tacaggtttg gattgaagta agggtggata caagatgact caattttggc tgaatcttcc
     7741 cttaaatcat caatacattt gccatcctac atgcctaatc aaatccagag gttttcagag
     7801 tttgggaaaa ctgctgattg catccttatc ccatccttcc tcttcctcac tgtgccgtat
     7861 agctgtgccc tgtctatcag ttcgggccat ggaagagcgt gtcacccagt cactcagggg
     7921 ctcagatagg cctggctccc tgctagtcct gggacgtggc ttatcaggca ggcctggctc
     7981 catttgaggg ggagtgggcc gacagcggct ccacagcagg cctgagcgcc accacccacc
     8041 atcccaaaaa aatactgata acacaaatca gcgagtgtgt gagacacagg agttcccatt
     8101 tcgcttatag tttccacaac aatgttaagc agcaaaccga aaagctgaca tattggggtg
     8161 atttctgaag gatcatgtgg cactgaagac tagagtaatg gtgctaaaaa tgcagatttg
     8221 cctgaatagc ttaaaaactg gcaatagttt aaggacaaat cctcactttt accagcctat
     8281 tattaagata ttggctgatt attggtagtt atgaagcaca tattctgcaa gatcttactc
     8341 tacatccata atcctcttca ataccgaaac ttaacctacg tatttaacat ttacattctc
     8401 actctgcgag cctgcttttt tgtacaaact tgtgatatcc tcgagggccc atctggcctg
     8461 tgtttcagac accagggagt ctctgctcac gtttcctgct atttgcagcc tc
//
