LOCUS       sequence-201487-        4107 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For expressing Trpa1b-2A-EGFP in zebrafish cardiomyocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4107)
  AUTHORS   Lam PY, Mendu SK, Mills RW, Zheng B, Padilla H, Milan DJ, Desai BN, 
            Peterson RT
  TITLE     A high-conductance chemo-optogenetic system based on the vertebrate 
            channel Trpa1b.
  JOURNAL   Sci Rep. 2017 Sep 19;7(1):11839. doi: 10.1038/s41598-017-11791-z.
  PUBMED    28928472
REFERENCE   2  (bases 1 to 4107)
  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 Rep."; 
            date: "2017-09-19"; volume: "7(1):11839. doi"; pages: " 
            10.1038/s41598-017-11791-z"
COMMENT     Sequence Label: Tol2-cmlc2-Trpa1b-2A-EGFP
FEATURES             Location/Qualifiers
     source          1..4107
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             3331..3387
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(3388..3390,3391..3393,3394..4107)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3388..3390,3391..3393,3394..4104)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3433..3454)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3694..3713)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4041..4062
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgcagtttg gaaaggagtt agtgaggcga aacagcttct acaaatgtgt gataaacgag
       61 gacgaaacgg aggagtctgc agatgtcttt gagttggctt tcaaaggaaa ggcttctgcc
      121 atcgatcgtc tgatccagaa aggtccagaa cacctaagcc tcagggatga gaatggagcc
      181 agtctgctgc actatgcttc agctggagga aacctggata tcattcggct cattgtttcc
      241 attgtgggcc ctgaggtgct gaatgtgcag gatgagcagg gccgaactcc tctgcactgg
      301 gctgtggagc aggatcaaca acagagctgc gctgtactgc tggatctggg tgctgacccc
      361 aacattctca acaatgccct gatgggccct ctgcacctgg ctgtcaccaa acaatacaat
      421 catctagcag aggtccttct gtcttgtgac aaaaccaata gcaatctgga gggggatctg
      481 ggaaacaccc ctgtcatgct ggcctgctcc aataacaact gccaagctct ccagatcctg
      541 atcaagcgcg gtgccaagat gtgtattcag aacaagctgg gtcactatcc cattcatact
      601 gttgctttcg caggtgccaa agaggccatg gagatggtat tgaaaattgg agaagaactt
      661 ggtgtatcat ctacattgca cattaattat ttggataagt ccaaaagcac acctcttcat
      721 ctggctgtac gcggaggcaa tatcgaggtg atcaaactct gcatcttgaa aggagccaaa
      781 gtagaacaac atcagagtgg caaatgcacg gccctccact ttgcctgcag tcagggttct
      841 ctggaggctg tcaaaatcat gctttcctct tacaacagaa cagaagatat cgtcaacatt
      901 cgggatggag ccaatcggac acctttgcat agagcaacgc tgtttgatca cgttgagttg
      961 gctgaatatc ttatttcaaa gggagcagaa attgactcta ttgactgtaa gggactgtct
     1021 ccgcttctgc tcgcctccag ctgcagcgct tggaaaacag ttgcatatct gttgtccatt
     1081 ggtgcagatt ttaaaatcaa agaaaaaaca ggtcgcaact tcctccactt tgtcattctt
     1141 caacccaaag gcctgaaaaa tcttcctgag acggttttgc agagcaccgc cgtgaaggag
     1201 atgttgtctg atgaggatgt tgaaggctgc actcctctgc actacgcctg caagcttggc
     1261 atccacgatt cggtcaaaaa tatgctgggt ctcaatatct gcttgggcca aaagtcacgg
     1321 gagaagaaat cagcgcttca cttcgctgca gaatatgggc gtataaatac atgccaccgg
     1381 ctgctggaga ccctcacaga ctctaaaatg ctgaatgatt gggatgaaaa gggtctgacg
     1441 cccctccatc ttgcttcaag ggcaggacac gctcaagtag tggatctgct gcttaggaag
     1501 ggagcactgt ttcaaagtga ttacaagggc tggacctgtt tacatcatgc agcagctgaa
     1561 ggatacacac agacgatgaa aatcctgcta gcagccaatg tgaaactact ggatgaaaaa
     1621 aatgaagatg ggaacacagc ccttcatata gcagcacaag ctggacatgt aagtgctgtt
     1681 ttgctgttat tggacagagg agctgaaatc gctctcaacg atgctgacaa ctcttttttg
     1741 catgaagctg tgcggaatga gagaagagaa gtagtaaacg ccaccattga acatgagcgg
     1801 tgtgatgagt caatgacgtc atttaaagcg aaatcccgct gtgttgtgct ggacatcata
     1861 gagttccttc cagaatcttt tcagcacctt ctggaccagt gtattacaga atctgatcat
     1921 gatgccaata gtcaggatta tcatatcatg tacaatttcc aatggcttca ggctcccata
     1981 cagttgaaga aatatgctaa aacagacaaa acaaaagcat ttcaaccact ggcagcatta
     2041 aatgccatgg tgcggtacaa ccgtctggag ctcctcattc accccttgag ccgaaaatac
     2101 ctggagatga agtggacggc atatggaagc aaagttcatt tcctaaactt ggcgatatac
     2161 ctgcttggat tgttgcccct cacatacctc atcctcaatc tgagacccag ccaggacttc
     2221 tccaaaggca atggcacatc tgtcatcatg gtgcccgtat cttttagcga gcaacaatat
     2281 ttaatctccg tgtgtataat catggtgatc gtcatgaatg tgtattcgat ttgcaaagag
     2341 gtggtacagc tggctcagca gcgagtaaat tacttcacag atttctcaaa ccctgcggac
     2401 tggagtgcag ctattagcgc tttggttttt gttgtaccca tgtgctgcag tgtggaggcc
     2461 acatggcaat gggaagctgg agcttatgct attctgacat catggattgg ctttctgctt
     2521 tactttcaga gatttgagcg gattgggatt tacgtggtga tgtttaatgg aatcgtacgg
     2581 accctggtgt gcatcatggt tctttttgtc ttccttttgc tggctttcgg cttggctttc
     2641 tacgctctga tgctccacag gccggagttc agttccatca gcctagcgtt agcgcaaacg
     2701 tttgtgatga cagtaggaga gctaaactac caaagcacat ttcttaactc ctatgaagaa
     2761 ggccacatgg ctttcccagc catcacttat ttagtttttg tgttttttgt tcttctcatg
     2821 cccattcttc tcatgaacct aatgattggt ttggctgtag gagacattgc agaggtacag
     2881 agaaacgcag agctgaaaag gattgctatg cagatagatc ttcacactgc ccttgaggag
     2941 aaactacctt actggttcct gaagcgagtg gacaaaccct ctaccgttgt ctatcccaat
     3001 aaatgcaaga aagtgcttct ggaggccttg gtgcatggtg agattcagaa cacagttcga
     3061 actcgtctaa acctttgctc acggaaggaa gggcttttgg agagagagct gcacaagcag
     3121 aagaacaggt tgaaagaaat gtcgtgcatg ctggaaaaac agcataacct gttgaagctg
     3181 attatccaga agatggaaat tacatctgaa gctgatgaat atgatggtcc acagaatcat
     3241 ggagctctaa aacagcacac ctcatccacc tcacagaaat caaagtgggt cccgttactg
     3301 caagcaatca aggccaagaa gggatccgga gccacgaact tctctctgtt aaagcaagca
     3361 ggagacgtgg aagaaaaccc cggtcctatg gtgagcaagg gcgaggagct gttcaccggg
     3421 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3481 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     3541 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     3601 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3661 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3721 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3781 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3841 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3901 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3961 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     4021 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     4081 ctcggcatgg acgagctgta caagtaa
//
