LOCUS       sequence-414325-        7987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitously expresses CoChR 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 7987)
  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 7987)
  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: pBJ016_MTB_CoChR_KGC_eGFP_ER2
FEATURES             Location/Qualifiers
     source          1..7987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    155..289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(179..198)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     233..252
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(381..518,519..530)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(589..606)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(589..605)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(598..620)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(764..783)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     974..995
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1158..1262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1263..1331,1332..2123)
                     /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(1481..1500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2294..2882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2783..2802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3036..3053
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3170..3191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3206..3211,3212..3229,3230..3236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3244..3260
                     /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     3249..3271
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3268..3284
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3268..3284
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3303..3323
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3305..3323
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3360..3376
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3360..3376
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(3410..3421,3422..3609)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     promoter        6325..6343
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6325..6342
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6351..6370
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6351..6369
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6407..7270
                     /codon_start=1
                     /product="high-photocurrent channelrhodopsin from 
                     Chloromonas oogama (Shemesh et al., 2017)"
                     /label=CoChR
                     /translation="MLGNGSAIVPIDQCFCLAWTDSLGSDTEQLVANILQWFAFGFSIL
                     ILMFYAYQTWRATCGWEEVYVCCVELTKVIIEFFHEFDDPSMLYLANGHRVQWLRYAEW
                     LLTCPVILIHLSNLTGLKDDYSKRTMRLLVSDVGTIVWGATSAMSTGYVKVIFFVLGCI
                     YGANTFFHAAKVYIESYHVVPKGRPRTVVRIMAWLFFLSWGMFPVLFVVGPEGFDAISV
                     YGSTIGHTIIDLMSKNCWGLLGHYLRVLIHQHIIIYGDIRKKTKINVAGEEMEVETMVD
                     QEDEETV"
     CDS             join(7379..7381,7382..7987,1..105)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
ORIGIN
        1 cagagcgctc tgagcaagga ccctaacgag aagagagacc acatggtgct gctggagttc
       61 gtgacagctg ctggaatcac actgggaatg gacgagctgt acaagttctg ttacgagaac
      121 gaggtgtgag aattcatacg tatccggaac cggtgatcca gacatgataa gatacattga
      181 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      241 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
      301 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt gatatcccgg
      361 gtttaaacgc atgcagatct aatactcaag tacaatttta atggagtact tttttacttt
      421 tactcaagta agattctagc cagatacttt tacttttaat tgagtaaaat tttccctaag
      481 tacttgtact ttcacttgag taaaattttt gagtactttt tacacctctg tcaagaacca
      541 tatgccggta cccaattcgc cctataggac gtcgtattac gcgcgctcac tggccgtcgt
      601 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
      661 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
      721 gttgcgcagc ctgaatggcg aatgggacgc gccctagcgc ccgctccttt cgctttcttc
      781 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
      841 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
      901 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
      961 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1021 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1081 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     1141 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1201 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1261 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1321 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1381 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1441 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1501 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1561 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1621 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1681 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1741 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1801 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1861 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1921 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     1981 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2041 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2101 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2161 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2221 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2281 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2341 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2401 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2461 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2521 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2581 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2641 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2701 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2761 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2821 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2881 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2941 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3001 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3061 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3121 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     3181 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     3241 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     3301 gcgcaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct
     3361 ctagaactag tggatctgct gggcttgctg aaggtagggg gtcaagaacc agaggtgtaa
     3421 agtacttgag taattttact tgattactgt acttaagtat tatttttggg gatttttact
     3481 ttacttgagt acaattaaaa atcaatactt ttacttttac ttaattacat ttttttagaa
     3541 aaaaaagtac tttttactcc ttacaatttt atttacagtc aaaaagtact tattttttgg
     3601 agatcacttg ggcccggctc gagggcctag tcattaccta tactcaacgt ttactctgag
     3661 aaacttccat attttagagg taaaatgctc agaattgtaa aaagtcttgt caaaatactt
     3721 ggaatttgcc aaaaacaaaa taaaagtcac atcacctcat tgtccttggt atcagcaagc
     3781 acttttaaca caatcagctg tttttgttta taactcccac tgaaggccat gtgcattact
     3841 cccaagttac acagtaaaaa gtcacatgta gtttatcatg atcactgctg ctcaagcaca
     3901 gctttatctg ttattcagtg tggcaatatc tgaaatgtgt gcaaaatatt atttattgta
     3961 gctgatcacc aacactttca gtgttttttt ttttctactt ttaatgagaa ttatgttctt
     4021 aggatttatc cttataaacc tctttaaaaa gaaatgttat gtctgcatag acacgacatg
     4081 aaattatgaa cagaaaattt aactatttaa gatcaagact tgacatataa catttctaat
     4141 gaacattaaa cctgaggtag gcccacttgc aaaaatgtaa ctgctatcac tcactttata
     4201 catcattgtg taacttgtac acacaaaaat agtgaagcaa tgtggcatgg aatgcaggtc
     4261 ccattcattc aaagttatta aagcactttc ttaaagtgca ttactactga gctaagacca
     4321 gctagttgct catacaacat actacaagta gtgtcatcct aatttggata aaaaaaatgt
     4381 ccattttata catggaaaag tactcattag atcccccatt gtttgtatta cggtatttcg
     4441 tgaacacaag aggtaaatga cctacagagc tgctgttgtg ttagattgaa aacacaacac
     4501 aggatcatgg aggcatccat cattatgacg cattctgcca cttgtaactc tgcacagact
     4561 ttttgaaaag tttaattgag tcttggcgtt gtcgcagaaa aatacgaaaa attctgtgtt
     4621 aaaatttgca aataaaaatt ccagttttat aggaaatcgc catggccttg ttctttaacc
     4681 agttaagctt ccccttcttt cactctcaag ttgcaagaag caagtgtagc aatgtgcacg
     4741 cgacagccgg gtgtgtgacg ctggaccaat cagagcacag agctccgaaa gtttaccttt
     4801 tatggctaga gccgggtatg tgccgtcata taaaagagcg cgcccagctt ttcagcctca
     4861 ctttgagctc ctccacacgc agctagtgcg gaatatcatc tgcttgtaac ccattctctt
     4921 aagtcgacaa ccccccaaac ccaaggtgag ttggttttac agtttttaag cgtttaatta
     4981 tagtttctat ctattaatgt taagtaaaat gtgtaaatgg ataaatatga tttgatgaag
     5041 tctttagaca gttcataacg gtctctttgt agttatctac taggtttgat ttataagtgg
     5101 tggttctttt gataatctat gttatattta ataaatattg gctgtatctt aacttttttg
     5161 tagcgttagt ggatgtggca ggtgagaatg tcggtaattt aacgtgacca gtcgtaggca
     5221 cgacattgaa cattgaatag gccggtgtga aatgggtgtt aagtcttaat ataaagttgt
     5281 gttctggcta atctctcttt aacagccata aatgtgattt ttaactacag ttttgagggt
     5341 gctggtgagc atgttgcaca cttgatggat agccggcatg ggaagttctt tgtgcaggca
     5401 gtgctgcagc agggtgtgac ctactttagc tagccggcta accagctctc atctgatgta
     5461 acgtaaaccc catcaatcta acatcggctc attcgtgctt tataagatcg aagatcttgt
     5521 tattagtaac gggtattggc atttagtttt aacttggtta attttctttt tgttatgtgt
     5581 tctatttaaa tgtagcctgc agccccctct gtctttaatg aagctgaaga ttaacgttaa
     5641 cgttagtgaa gtgttgtatc cgctatgaaa gatttgaatt ttagaggtgt ggtccacttt
     5701 aataacggct gcggagttct ctggctgcac ttcagccgct gtgactgtcg aagcagagcc
     5761 catcagtttt tcaggatctc ggtgattttt gaggctagac tatgaactga accgactgag
     5821 agtgaccgtc cagaaatgct ttagtgtgaa gtctccgctc acgaggagtc cgtggactga
     5881 ctgcagatct gtgacgcagt aacacccggc ggcagacacc cgttgaaatg cggttgtgta
     5941 attgatcgca ggcgagggtg gaagaggatg tgaaacttca tttgtgtaaa atttagggag
     6001 tggtcctggt gtgatgaatg tcgaaatctg ttccttttta ctgagcccta cgactctggc
     6061 tgagtgccac accgccggca gccgcaaagc ggctcaaacc attgcctttt atggtaataa
     6121 tgagagaatg cagagggact tcctttgtct ggcatatctg aggcgcgcgc tgtcactagc
     6181 gcccaccagc ggtcagactg tagaatgcag agcaaaacag gaagttgact ccagatggtc
     6241 acatgcttgc tgaaacgcct ctccctggca gcagtgcaat tctaacagtt tccctttctt
     6301 ttacagttca gccatcctag gcctatttag gtgacactat agaagagtac taatacgact
     6361 cactataggg agaagctact tgttcttttt gcaggatccc gccaccatgc tgggaaacgg
     6421 cagcgccatt gtgcctatcg accagtgctt ttgcctggct tggaccgaca gcctgggaag
     6481 cgatacagag cagctggtgg ccaacatcct ccagtggttc gccttcggct tcagcatcct
     6541 gatcctgatg ttctacgcct accagacttg gagagccact tgcggttggg aggaggtcta
     6601 cgtctgttgc gtcgagctga ccaaggtcat catcgagttc ttccacgagt tcgacgaccc
     6661 cagcatgctg tacctggcta acggacaccg agtccagtgg ctgagatacg cagagtggct
     6721 gctgacttgt cccgtcatcc tgatccacct gagcaacctg accggcctga aggacgacta
     6781 cagcaagcgg accatgaggc tgctggtgtc agacgtggga accatcgtgt ggggagctac
     6841 aagcgccatg agcacaggct acgtcaaggt catcttcttc gtgctgggtt gcatctacgg
     6901 cgccaacacc ttcttccacg ccgccaaggt gtatatcgag agctaccacg tggtgccaaa
     6961 gggcagacct agaaccgtcg tgcggatcat ggcttggctg ttcttcctgt cttggggcat
     7021 gttccccgtg ctgttcgtcg tgggaccaga aggattcgac gccatcagcg tgtacggctc
     7081 taccattggc cacaccatca tcgacctcat gagcaagaat tgttggggcc tgctgggaca
     7141 ctatctgaga gtgctgatcc accagcacat catcatctac ggcgacatcc ggaagaagac
     7201 caagatcaac gtggccggcg aggagatgga agtggagacc atggtggacc aggaggacga
     7261 ggagacagtg agatctgctg ctgacagacc tgtggtggct gtgagcaagg ctgctgctaa
     7321 gagcagaatc acaagcgagg gagagtacat ccctctggac cagatcgaca tcaacgtggt
     7381 gagcaaggga gaggagctgt tcacaggagt ggtgcctatc ctggtggagc tggacggaga
     7441 cgtgaacgga cacaagttca gcgtgagcgg agagggagag ggagacgcta catacggaaa
     7501 gctgacactg aagttcatct gtacaacagg aaagctgcct gtgccttggc ctacactggt
     7561 gacaacactg acatacggag tgcagtgttt cagcagatac cctgaccaca tgaagcagca
     7621 cgacttcttc aagagcgcta tgcctgaggg atacgtgcag gagagaacaa tcttcttcaa
     7681 ggacgacgga aactacaaga caagagctga ggtgaagttc gagggagaca cactggtgaa
     7741 cagaatcgag ctgaagggaa tcgacttcaa ggaggacgga aacatcctgg gacacaagct
     7801 ggagtacaac tacaacagcc acaacgtgta catcatggct gacaagcaga agaacggaat
     7861 caaggtgaac ttcaagatca gacacaacat cgaggacgga agcgtgcagc tggctgacca
     7921 ctaccagcag aacacaccta tcggagacgg acctgtgctg ctgcctgaca accactacct
     7981 gagcaca
//
