LOCUS       sequence-414333-        7519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitously expresses FR-GECO1c in zebrafish via tol2 transgenesis 
            or in vitro transcription of mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7519)
  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 7519)
  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: pBJ037_MTB_FR-GECO1c
FEATURES             Location/Qualifiers
     source          1..7519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     367..384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    501..522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(537..542,543..560,561..567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    575..591
                     /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     580..602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     599..615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     599..615
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     634..654
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        636..654
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     691..707
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     691..707
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(741..752,753..940)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        3656..3674
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3656..3673
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     3682..3701
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3682..3700
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3732..3741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5005..5139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5029..5048)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5083..5102
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5231..5368,5369..5380)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5439..5456)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5439..5455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5448..5470)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5614..5633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5824..5845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6008..6112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6113..6181,6182..6973)
                     /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(6331..6350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7144..7519,1..213)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
       61 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
      421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
      481 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      541 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      601 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
      661 aagctggagc tccaccgcgg tggcggccgc tctagaacta gtggatctgc tgggcttgct
      721 gaaggtaggg ggtcaagaac cagaggtgta aagtacttga gtaattttac ttgattactg
      781 tacttaagta ttatttttgg ggatttttac tttacttgag tacaattaaa aatcaatact
      841 tttactttta cttaattaca tttttttaga aaaaaaagta ctttttactc cttacaattt
      901 tatttacagt caaaaagtac ttattttttg gagatcactt gggcccggct cgagggccta
      961 gtcattacct atactcaacg tttactctga gaaacttcca tattttagag gtaaaatgct
     1021 cagaattgta aaaagtcttg tcaaaatact tggaatttgc caaaaacaaa ataaaagtca
     1081 catcacctca ttgtccttgg tatcagcaag cacttttaac acaatcagct gtttttgttt
     1141 ataactccca ctgaaggcca tgtgcattac tcccaagtta cacagtaaaa agtcacatgt
     1201 agtttatcat gatcactgct gctcaagcac agctttatct gttattcagt gtggcaatat
     1261 ctgaaatgtg tgcaaaatat tatttattgt agctgatcac caacactttc agtgtttttt
     1321 tttttctact tttaatgaga attatgttct taggatttat ccttataaac ctctttaaaa
     1381 agaaatgtta tgtctgcata gacacgacat gaaattatga acagaaaatt taactattta
     1441 agatcaagac ttgacatata acatttctaa tgaacattaa acctgaggta ggcccacttg
     1501 caaaaatgta actgctatca ctcactttat acatcattgt gtaacttgta cacacaaaaa
     1561 tagtgaagca atgtggcatg gaatgcaggt cccattcatt caaagttatt aaagcacttt
     1621 cttaaagtgc attactactg agctaagacc agctagttgc tcatacaaca tactacaagt
     1681 agtgtcatcc taatttggat aaaaaaaatg tccattttat acatggaaaa gtactcatta
     1741 gatcccccat tgtttgtatt acggtatttc gtgaacacaa gaggtaaatg acctacagag
     1801 ctgctgttgt gttagattga aaacacaaca caggatcatg gaggcatcca tcattatgac
     1861 gcattctgcc acttgtaact ctgcacagac tttttgaaaa gtttaattga gtcttggcgt
     1921 tgtcgcagaa aaatacgaaa aattctgtgt taaaatttgc aaataaaaat tccagtttta
     1981 taggaaatcg ccatggcctt gttctttaac cagttaagct tccccttctt tcactctcaa
     2041 gttgcaagaa gcaagtgtag caatgtgcac gcgacagccg ggtgtgtgac gctggaccaa
     2101 tcagagcaca gagctccgaa agtttacctt ttatggctag agccgggtat gtgccgtcat
     2161 ataaaagagc gcgcccagct tttcagcctc actttgagct cctccacacg cagctagtgc
     2221 ggaatatcat ctgcttgtaa cccattctct taagtcgaca accccccaaa cccaaggtga
     2281 gttggtttta cagtttttaa gcgtttaatt atagtttcta tctattaatg ttaagtaaaa
     2341 tgtgtaaatg gataaatatg atttgatgaa gtctttagac agttcataac ggtctctttg
     2401 tagttatcta ctaggtttga tttataagtg gtggttcttt tgataatcta tgttatattt
     2461 aataaatatt ggctgtatct taactttttt gtagcgttag tggatgtggc aggtgagaat
     2521 gtcggtaatt taacgtgacc agtcgtaggc acgacattga acattgaata ggccggtgtg
     2581 aaatgggtgt taagtcttaa tataaagttg tgttctggct aatctctctt taacagccat
     2641 aaatgtgatt tttaactaca gttttgaggg tgctggtgag catgttgcac acttgatgga
     2701 tagccggcat gggaagttct ttgtgcaggc agtgctgcag cagggtgtga cctactttag
     2761 ctagccggct aaccagctct catctgatgt aacgtaaacc ccatcaatct aacatcggct
     2821 cattcgtgct ttataagatc gaagatcttg ttattagtaa cgggtattgg catttagttt
     2881 taacttggtt aattttcttt ttgttatgtg ttctatttaa atgtagcctg cagccccctc
     2941 tgtctttaat gaagctgaag attaacgtta acgttagtga agtgttgtat ccgctatgaa
     3001 agatttgaat tttagaggtg tggtccactt taataacggc tgcggagttc tctggctgca
     3061 cttcagccgc tgtgactgtc gaagcagagc ccatcagttt ttcaggatct cggtgatttt
     3121 tgaggctaga ctatgaactg aaccgactga gagtgaccgt ccagaaatgc tttagtgtga
     3181 agtctccgct cacgaggagt ccgtggactg actgcagatc tgtgacgcag taacacccgg
     3241 cggcagacac ccgttgaaat gcggttgtgt aattgatcgc aggcgagggt ggaagaggat
     3301 gtgaaacttc atttgtgtaa aatttaggga gtggtcctgg tgtgatgaat gtcgaaatct
     3361 gttccttttt actgagccct acgactctgg ctgagtgcca caccgccggc agccgcaaag
     3421 cggctcaaac cattgccttt tatggtaata atgagagaat gcagagggac ttcctttgtc
     3481 tggcatatct gaggcgcgcg ctgtcactag cgcccaccag cggtcagact gtagaatgca
     3541 gagcaaaaca ggaagttgac tccagatggt cacatgcttg ctgaaacgcc tctccctggc
     3601 agcagtgcaa ttctaacagt ttccctttct tttacagttc agccatccta ggcctattta
     3661 ggtgacacta tagaagagta ctaatacgac tcactatagg gagaagctac ttgttctttt
     3721 tgcaggatcc cgccaccatg ggcagcgtga agcagatccc tagcctgacc acagtgatcc
     3781 tggtcaagtc catgctgcgg aagcggagct tcggcaaccc cttcggtttt ctgaccgaga
     3841 ccctggtgcc tgtggacggt ggcctggcag gcacatgtga catggccctg agactcgttg
     3901 gcggcggaca cctgaagtgc aacctgaaaa ccacctacag aagcaagaag cccgccaaga
     3961 acctgaagat gcctggcgaa caccaggtgg gccggaagct ggaaagaatc aaagaggccg
     4021 acaacgagac atacgtggaa cagcacgaag tggccgtggc cagatatgtt ggacttggcg
     4081 gaggcggagg aacaggcgga tctgtgtctg agctgatcaa agaaaacatg cggatgaagc
     4141 tctacttgga agggaccgtc ggcaaccacc acttcaagtg tactgccgaa ggcgagggca
     4201 agccctatga gggaacacag acccagcgga tcaaggtggt ggaaggtggc cctctgcctt
     4261 tcgcctttga catcctggcc acctgtttta tgtacggcag caagaccttc atcaactatc
     4321 cccagggcat ccccgatttc ttcaagcaga gcttccccga gggcttcacc tgggagagag
     4381 tgaccacata cgaggatggc ggcgtgctga ccgtgacaca ggatacaagc ctccaggacg
     4441 gctacctgat ctacaatgtg aagctgctgg gcgtgaactt ccccagcgat ggacccgtga
     4501 tgcagaagaa aaccctcggc tgggtgccca ccgatggcac gctgacagaa gaactgatcg
     4561 ccgagttcaa agaagccttc agcctgttcg acaaggacgg cgacggcacc atcaccacca
     4621 aagaactggg ctccgtgatg cggagcctgg gccagaatcc aacagaagcc gaactccagg
     4681 acatgatcaa cgaggtggac gccgatggcg acggaacctt cgattaccct gagttcctga
     4741 ccatgatgtc ccggaagatg agctacagag tgaccgagga agagatccgg gaagccttcc
     4801 aggttttcga taaggatggc aacggctaca tcggagccgc cgaactgaga cgcgtgatgg
     4861 ccgatctggg cgagaagctg actgacgaag aggttgacga gatgatcaga gtggccgaca
     4921 tcgacggcga cggacaagtg aactacgaag agttcgtgca gatgatgacc gccaagtagg
     4981 aattcatacg tatccggaac cggtgatcca gacatgataa gatacattga tgagtttgga
     5041 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     5101 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     5161 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt gatatcccgg gtttaaacgc
     5221 atgcagatct aatactcaag tacaatttta atggagtact tttttacttt tactcaagta
     5281 agattctagc cagatacttt tacttttaat tgagtaaaat tttccctaag tacttgtact
     5341 ttcacttgag taaaattttt gagtactttt tacacctctg tcaagaacca tatgccggta
     5401 cccaattcgc cctataggac gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     5461 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5521 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5581 ctgaatggcg aatgggacgc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5641 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5701 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5761 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5821 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5881 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5941 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     6001 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6121 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6181 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6241 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6301 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6361 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6421 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6481 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6541 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6601 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6661 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6721 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6781 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6841 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6901 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6961 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7021 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7081 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7141 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7201 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7261 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7321 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7381 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7441 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7501 gacctacacc gaactgaga
//
