LOCUS       sequence-207459-        7112 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For expressing Trpa1b-2A-mCherry in zebrafish Rohon-Beard neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7112)
  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 7112)
  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-ngn1-Trpa1b-2A-mCherry
FEATURES             Location/Qualifiers
     source          1..7112
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(730..748)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    748..772
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4110..4166
                     /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             4167..4877
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4312..4330)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4580..4599
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(4879..4903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4905..5039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(5102..5122)
                     /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(5130..5146)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5162..5296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(5735..5922,5923..5934)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5968..5984)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5968..5984)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(6021..6039)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6060..6076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6060..6076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6073..6095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6084..6100
                     /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(6108..6114,6115..6132,6133..6138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6153..6174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6291..6308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6462..7050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6542..6561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catagaaaaa tatttataac atatataaca tttagaattc tgcaacaaac caaaacaatt
       61 tggccttaaa taatatttga cagatatgat gtgatataac aaaatacatt agaaaaatgt
      121 gcaaaatgtg tcttgcagcc tcaggccact tttgctgaag tccattcttt tcgatttcct
      181 cttgagattt aagttaaagt gggcttgttg catgcaggac agacagcaac agctccgctc
      241 gcgtggccgg acagttctcc tgctctctga ttggccagcg gtgattacag gcacgcttca
      301 gagggcgaac aaacaacagg cataaattat gctaatgaga tggtcgtgcg gcacggggca
      361 gacctgatgc aaaccattca ttcatccttc acaaaaatgc caaagcgagg ggggtatgtc
      421 cattgtccca tcagggtata taaggtaggt gaggccagtc gtgtgccaag ctcacaagct
      481 cacacgagct gattgagggt taagagcaag acagacgcgt gccattatct tcaacacaaa
      541 ctatctgcag gattctgcaa aacctcaagc atctcccagc ccaccaataa ggtgcgtatt
      601 gggtcacgca gataacttga aaggactttg cacgactgat gttgtttttt gtttcttttc
      661 tggcgtattc ctaacctgat tttgtaaatt tctccaggtt atcaaccggt ggagctccag
      721 cttttgttcc ctttagtgag ggttaattca agtttgtaca aaaaagcggc ttcgccacca
      781 tgcagtttgg aaaggagtta gtgaggcgaa acagcttcta caaatgtgtg ataaacgagg
      841 acgaaacgga ggagtctgca gatgtctttg agttggcttt caaaggaaag gcttctgcca
      901 tcgatcgtct gatccagaaa ggtccagaac acctaagcct cagggatgag aatggagcca
      961 gtctgctgca ctatgcttca gctggaggaa acctggatat cattcggctc attgtttcca
     1021 ttgtgggccc tgaggtgctg aatgtgcagg atgagcaggg ccgaactcct ctgcactggg
     1081 ctgtggagca ggatcaacaa cagagctgcg ctgtactgct ggatctgggt gctgacccca
     1141 acattctcaa caatgccctg atgggccctc tgcacctggc tgtcaccaaa caatacaatc
     1201 atctagcaga ggtccttctg tcttgtgaca aaaccaatag caatctggag ggggatctgg
     1261 gaaacacccc tgtcatgctg gcctgctcca ataacaactg ccaagctctc cagatcctga
     1321 tcaagcgcgg tgccaagatg tgtattcaga acaagctggg tcactatccc attcatactg
     1381 ttgctttcgc aggtgccaaa gaggccatgg agatggtatt gaaaattgga gaagaacttg
     1441 gtgtatcatc tacattgcac attaattatt tggataagtc caaaagcaca cctcttcatc
     1501 tggctgtacg cggaggcaat atcgaggtga tcaaactctg catcttgaaa ggagccaaag
     1561 tagaacaaca tcagagtggc aaatgcacgg ccctccactt tgcctgcagt cagggttctc
     1621 tggaggctgt caaaatcatg ctttcctctt acaacagaac agaagatatc gtcaacattc
     1681 gggatggagc caatcggaca cctttgcata gagcaacgct gtttgatcac gttgagttgg
     1741 ctgaatatct tatttcaaag ggagcagaaa ttgactctat tgactgtaag ggactgtctc
     1801 cgcttctgct cgcctccagc tgcagcgctt ggaaaacagt tgcatatctg ttgtccattg
     1861 gtgcagattt taaaatcaaa gaaaaaacag gtcgcaactt cctccacttt gtcattcttc
     1921 aacccaaagg cctgaaaaat cttcctgaga cggttttgca gagcaccgcc gtgaaggaga
     1981 tgttgtctga tgaggatgtt gaaggctgca ctcctctgca ctacgcctgc aagcttggca
     2041 tccacgattc ggtcaaaaat atgctgggtc tcaatatctg cttgggccaa aagtcacggg
     2101 agaagaaatc agcgcttcac ttcgctgcag aatatgggcg tataaataca tgccaccggc
     2161 tgctggagac cctcacagac tctaaaatgc tgaatgattg ggatgaaaag ggtctgacgc
     2221 ccctccatct tgcttcaagg gcaggacacg ctcaagtagt ggatctgctg cttaggaagg
     2281 gagcactgtt tcaaagtgat tacaagggct ggacctgttt acatcatgca gcagctgaag
     2341 gatacacaca gacgatgaaa atcctgctag cagccaatgt gaaactactg gatgaaaaaa
     2401 atgaagatgg gaacacagcc cttcatatag cagcacaagc tggacatgta agtgctgttt
     2461 tgctgttatt ggacagagga gctgaaatcg ctctcaacga tgctgacaac tcttttttgc
     2521 atgaagctgt gcggaatgag agaagagaag tagtaaacgc caccattgaa catgagcggt
     2581 gtgatgagtc aatgacgtca tttaaagcga aatcccgctg tgttgtgctg gacatcatag
     2641 agttccttcc agaatctttt cagcaccttc tggaccagtg tattacagaa tctgatcatg
     2701 atgccaatag tcaggattat catatcatgt acaatttcca atggcttcag gctcccatac
     2761 agttgaagaa atatgctaaa acagacaaaa caaaagcatt tcaaccactg gcagcattaa
     2821 atgccatggt gcggtacaac cgtctggagc tcctcattca ccccttgagc cgaaaatacc
     2881 tggagatgaa gtggacggca tatggaagca aagttcattt cctaaacttg gcgatatacc
     2941 tgcttggatt gttgcccctc acatacctca tcctcaatct gagacccagc caggacttct
     3001 ccaaaggcaa tggcacatct gtcatcatgg tgcccgtatc ttttagcgag caacaatatt
     3061 taatctccgt gtgtataatc atggtgatcg tcatgaatgt gtattcgatt tgcaaagagg
     3121 tggtacagct ggctcagcag cgagtaaatt acttcacaga tttctcaaac cctgcggact
     3181 ggagtgcagc tattagcgct ttggtttttg ttgtacccat gtgctgcagt gtggaggcca
     3241 catggcaatg ggaagctgga gcttatgcta ttctgacatc atggattggc tttctgcttt
     3301 actttcagag atttgagcgg attgggattt acgtggtgat gtttaatgga atcgtacgga
     3361 ccctggtgtg catcatggtt ctttttgtct tccttttgct ggctttcggc ttggctttct
     3421 acgctctgat gctccacagg ccggagttca gttccatcag cctagcgtta gcgcaaacgt
     3481 ttgtgatgac agtaggagag ctaaactacc aaagcacatt tcttaactcc tatgaagaag
     3541 gccacatggc tttcccagcc atcacttatt tagtttttgt gttttttgtt cttctcatgc
     3601 ccattcttct catgaaccta atgattggtt tggctgtagg agacattgca gaggtacaga
     3661 gaaacgcaga gctgaaaagg attgctatgc agatagatct tcacactgcc cttgaggaga
     3721 aactacctta ctggttcctg aagcgagtgg acaaaccctc taccgttgtc tatcccaata
     3781 aatgcaagaa agtgcttctg gaggccttgg tgcatggtga gattcagaac acagttcgaa
     3841 ctcgtctaaa cctttgctca cggaaggaag ggcttttgga gagagagctg cacaagcaga
     3901 agaacaggtt gaaagaaatg tcgtgcatgc tggaaaaaca gcataacctg ttgaagctga
     3961 ttatccagaa gatggaaatt acatctgaag ctgatgaata tgatggtcca cagaatcatg
     4021 gagctctaaa acagcacacc tcatccacct cacagaaatc aaagtgggtc ccgttactgc
     4081 aagcaatcaa ggccaagaag ggatccggag ccacgaactt ctctctgtta aagcaagcag
     4141 gagacgtgga agaaaacccc ggtcctatgg tgagcaaggg cgaggaggat aacatggcca
     4201 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     4261 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     4321 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     4381 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     4441 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     4501 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     4561 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     4621 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     4681 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     4741 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     4801 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     4861 acgagctgta caagtaagac ccagctttct tgtacaaagt gggggatcca gacatgataa
     4921 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4981 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     5041 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     5101 ccaactttat tatacatagt tgataattca ctggccgtcg ttttacggta ccatcgatga
     5161 tgatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     5221 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     5281 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     5341 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     5401 tcctctagat cagatctgcg aagatacggc cacgggtgct cttgatcctg tggctgattt
     5461 tggactgtgc tgctcgcagc tgctgatgaa tcacatactt cctccatttt cttccactga
     5521 ttgactgtta taatttccct aatttccagg tcaaggtgct gtgcattgtg gtaatagatg
     5581 tgacatgacg tcacttccaa aggaccaatg aacatgtctg accaatttca tataatgtga
     5641 aaacgatttt cataggcaga ataaataaca tttaaattaa actgggcatc agcgcaattc
     5701 aattggtttg gtaatagcaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt
     5761 tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat
     5821 taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa
     5881 ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctggttctt
     5941 gaccccctac cttcagcaag cccagcagat ccactagttc tagagcggcc gccaccgcgg
     6001 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     6061 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6121 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6181 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6241 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6301 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6361 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6421 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6481 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6541 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6601 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6661 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6721 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6781 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6841 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6901 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6961 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7021 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7081 gacgctcagt ggaacgaaaa ctcacgttaa gg
//
