LOCUS       sequence-332390-        9783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of YFP-CaCyclOp(A-2x) in cholinergic motor neurons of C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9783)
  AUTHORS   Henss T, Nagpal J, Gao S, Scheib U, Pieragnolo A, Hirschhauser A, 
            Schneider-Warme F, Hegemann P, Nagel G, Gottschalk A
  TITLE     Optogenetic tools for manipulation of cyclic nucleotides 
            functionally coupled to cyclic nucleotide-gated channels.
  JOURNAL   Br J Pharmacol. 2021 Mar 18. doi: 10.1111/bph.15445.
  PUBMED    33733470
REFERENCE   2  (bases 1 to 9783)
  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: "Br J 
            Pharmacol. 2021 Mar 18. doi: 10.1111/bph.15445."
COMMENT     Sequence Label: pTH42 [punc-17::YFP-CaCyclOp(A-2x)::SL2::mCherry]
FEATURES             Location/Qualifiers
     source          1..9783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(3240..3242,3243..3245,3246..3956)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3285..3306)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3546..3565)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3893..3914
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6115..6825
                     /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(6260..6278)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6528..6547
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7741..7763)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7863..7882
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7964..7982)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8050..8154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8155..8223,8224..9015)
                     /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(8373..8392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9186..9774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9675..9694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gggattacac
       61 caatcatttc tcccctccga aatgtgtagt cgtcatgaca aagtggtgac actggattac
      121 ggtaggctcc gcccacttct tggaaaagtg acgtgggctc ggtgagccct aaagcttttg
      181 gttttcacaa ttttctggtt tttgggattt ttgcgggaat tttgagagga gaaagttggg
      241 agagaagata ttggaatggt ccccggggag ctgtacattt atacatctag actgaagctg
      301 gggtattgca aaaagagaaa agcctatcac tttctttcca tcctgcatac ttgccaaaaa
      361 gttttggctg agcttatttt taaaattgga attttttgga attttcagaa aagaaaaagt
      421 tttttctttt tttttaattt aaaaaatttt tgctatttat tgcatggtac tacagaaaaa
      481 cttgaatgaa ataaaaaatt tggaaaattc aagaaaactt tttttttcat agtttattta
      541 ttcgagtttt aatgtggctt ttattttgtt cggtttttgt tttaaaaaga ttttatacaa
      601 aatttcaaga atttacaatt tttccgttgt aaaaaatcca aaaaaaaatc caaaaaaata
      661 gttcagacct ttttttaaat tttaaattta aaaattcaat accaaaaaat taaaaaaaat
      721 tagtttttcc gagaataaag gaaaaatgtt gaaaaaatat tatatataaa atttccagaa
      781 aatgtgaaaa ctccaaaaaa ctgcagactt ttccccaaac tagctatttt ttgaattcat
      841 tacctttaaa ggtggagtac cgaaatttaa gactttgctt ttttagaccc aaaatggtcc
      901 aaaactacca aatttcgtaa tgagacgttc tgaaaatttt tcaaaaaaat gttatgggag
      961 ctcaaagttt ttgaaaaaat ggtatatttt tagctaattt taaccacttc ttggcgtcgc
     1021 agcggttgga actcaatttt aattaaatta tcatccttca atgtccttca atctttgcaa
     1081 tttcaatcga ccatacgcgt tcagagcatt tttttttttc gtttttcctc ccgtccagca
     1141 catatttaaa tattctctct ccccctcccc tggttcctaa aaacttggga aaggggaatt
     1201 tcgaaggagg tggaagcttc ttcttctccc cgagcacaca cacacatcct ctcttcaaaa
     1261 tctttaagaa atagtgcgtg tgaatgacta gacgctgaag ctaacttggt ttccgcgaac
     1321 aatcccaatc agtgcctgtg ccgaggcttc ggagaagaag agattgacgt cgtctttttc
     1381 tttttttgtg aaacagaata aaaaagaata cgaaatggac tttttcacac attttcgtta
     1441 cttttccaga aaaaaacgaa gctctagcca tggaagtttg aattgaaaac ctaaaccccg
     1501 atgtgtattc atttgagctt aagcctaagc ctaattctac gcctaagcat aagcctaagc
     1561 ctatgttcaa gcctaagcca aattgtagaa cgttaccaaa agatcaaacc tatttttcct
     1621 acagaaaagg tctaatgata actgttaagc gagagacaag gagagatata tatcaaaaac
     1681 tagcccaaac tgtgattaac atctattaac tgacctcaaa aaaaaaaata gtagagatta
     1741 aatactgctc aaaaagtttc ttggaaaagt aaattgttag gtttctgggg cctgtaaaac
     1801 tggaccgagt tttgaagaat ttttttaact tttcaaagtt tggcgtcact gaaagcttta
     1861 tattgtacat gttccaccaa ttttcttttg tatccgtttt ttctgtctca aaacggagct
     1921 cgagcccatg acgtgtggat accgtgagaa tccgcccaat tccttcacat cgatctcata
     1981 tttctttccc agtttatatt gaattcccgt tgaattgcgg ctttgcaggg gagaagatgt
     2041 gcgtctttat attttttttt ctatccttac ccgccgtgaa tccggctcga attgagccaa
     2101 tttttatgaa tagtagagct aattgacttc gtctccctag aattgatgag gtaaagctcg
     2161 tggggagata tggagatttt tcgatttttt ttccagtaat tttcagttca aaatttgagc
     2221 ttttgagcag gagttttgag atttgtgcca aaatttccta ttattttttc aaatattttt
     2281 gcttttgtaa aaaaggtgta gtagaagttt tgtagtgcca ctattttttt ttgaaaatta
     2341 tgttttggcg gaaagttctt taaaaaagtt aatcaaaatt tttggcgctt tagtcaaatt
     2401 aattttctgg caaaaatatt agtaattttg gcgtgagttc aattttccca ttcttaaaat
     2461 ttcacacaat taagaatttt aagatttggg tttttttttc aacaaaaaat caaatattta
     2521 catgaaaatt cggcgggaaa taaaaaaaaa tattccgatt caatgtgtga ataacttttt
     2581 gcggggaatt caaatttgca ttatttaaac tttttttaag ctcgaaaaat tgaaattctt
     2641 aaataaataa ataaaatttt cctcgaattt tgaatgtttc gattttttag cgttattttt
     2701 tcaaaatttc cactaaaact ctagttctaa agttctaaaa ctttaaaaac ttcctaatgt
     2761 tgtggtcttg tgtttttttg gcagcgattt ccaaatttct gaaattccaa caaaaaaacc
     2821 actacgtcag tttgcatatc caaccccccc cccccccccc tctgagccaa tcaatcatgt
     2881 gcattttgaa ttggcgccaa ggcaatcttc gtaaaaaata acaatagagt catccctcag
     2941 aacagaaaag aaacccgccc gggggctcaa atacacatgt catttttctt ttttcgcctg
     3001 atttttagcc taaattagtg tttagcaatt ttttaaggta actcggtgga gtcatttgaa
     3061 ctttttaaac taaaatttca gaaaaaattc tttttttttg tcaaaaaata attttcccgc
     3121 cggaattttt caaaaaaata atttttattt tatttttcaa gttccttgtt tcctctttat
     3181 ttttttattt tgcccccaaa agttttacac taaaatattt aattttcagc tcggctagca
     3241 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3301 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3361 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3421 tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     3481 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3541 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3601 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3661 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     3721 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     3781 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3841 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3901 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggat
     3961 caatgtctat gaaagataaa gataataacc tgcggggagc atgtagtggc tgttcctgcc
     4021 ctgaatactg ttactcacca acctcaaccc tgtgcgacga ttgcaagtgt agcgtgacaa
     4081 aacaccctat cgtcgagcag ccactgactc gaaacggaag tttccggagc tccggagcct
     4141 cactgctgcc atctcccagt cagcccaata tcaaggtcac tggctctagt accgcctcaa
     4201 gcaacgctaa tatgcggaac agacagaaca attcactgag cgtgtccaat gtccgcagca
     4261 cctcctctgc aagttcaagc aacgtgtcct ctcccgccaa tagccgacct ggctctccta
     4321 gtaagcagtc cgctctgcag cagtaccaga caaacatcgc agacatgtgg tcctgggata
     4381 tgatgctgag taccccttca ctgaaattcc tgacaggaca gtttatcatg tgggccattc
     4441 tgaccgtggc cggcgctttc tacgctctgt ttatccagga aagacaggct tataataggg
     4501 gctgggcaga catttggtac ggctatgggg ccttcggatt tggcatcggg attgctttca
     4561 gttatatggg atttgcaggc gccagaaacc cagagaagaa agcactgtcc ctgtgcctgc
     4621 tgggagtgaa catcatcgct tttagttcat acatcctgat tatgctgcgg ctgactccaa
     4681 ccatcgaagg caccctgagc aacccagtgg agcctgcaag atacctggag tggattgcta
     4741 cctgtcccgt cctgattctg ctgatctccg agattacaca ggcagaccat aatgcctggg
     4801 gggtggtctt ctctgattac gccctggtgg tctgcgggtt ctttggagct gtgctgcccc
     4861 cttatccctg gggaaacctg tttaatatcc tgtcatgtgc cttctttagc ttcgtggtct
     4921 acagcctgtg gaggtccttt actggggcca tcaacggaga gaccccttgc aacattgaag
     4981 tgaatggcct gaggtggaca cgcttctcaa ctgtcaccac atggaccctg tttccactga
     5041 gctggttcgc ctttacatct ggcatgctga gtttcacaat gactgaggca tcttttacca
     5101 tgatcgatat tggggccaag gtgttcctga cactggtgct ggtcaacagc actgtggagc
     5161 aggcacagaa tcagaaagtc gaagccatca cagctattgc agaggaactg gaatctcaga
     5221 tcactaactg tgacgccatt ctgcagaaga tgatgcctga gggggtgctg gaacagctga
     5281 agaatggaca ggcaaccgag gccaaagagt acgaatccgt gacagtcttc ttttctgaca
     5341 tcacaaactt tactgtgatt agctccagga cctccacaaa ggacatgatg gccaccctga
     5401 acaagctgtg gctggagtat gatgccatcg ctaagcggtg gggcgtgtat aaggtcaaga
     5461 ctattgggga cgcttatctg ggagtgaccg gagcaccaga ggtggtcccc gaccacgctg
     5521 atcgagcagt gaacttcgcc ctggacatca tcgagatgat caagactttt aaaactgcta
     5581 ccggcgaatc tatcaacatt cgcatcggcc tgaatagtgg accagtgaca gccggagtcc
     5641 tgggcgacct gaaccctcat tgggatctgg tgggggatac cgtcaataca gcttcccgga
     5701 tggagtcaac cagcaaggca gggcacattc atatctccga ctctacttac cagatgatca
     5761 agggaaaatt cgtgacccag ccactggacc tgatggaagt gaaaggaaag ggaaaaatgc
     5821 agacatattg ggtgaccgct agaaagtaag gtaccgctgt ctcatcctac tttcacctag
     5881 ttaactgctt gtcttaaaat ctatgcttct ctttagtatc taaaattttc ctagaagctt
     5941 acaagtatat aaatggtctc ttctcaataa aggttgtata tttattcatc ttattgaatc
     6001 tgccatttcc tcgtttttgc gagtttatat accttccaat tttctttcta ttgtattttc
     6061 aacttctaat tttaattcag ggaaactgct gtaccggtag aaaaaatgct cgagatggtg
     6121 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     6181 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     6241 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     6301 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     6361 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     6421 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     6481 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     6541 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     6601 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     6661 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     6721 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     6781 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagaatt ccaactgagc
     6841 gccggtcgct accattacca acttgtctgg tgtcaaaaat aataggggcc gctgtcatca
     6901 gagtaagttt aaactgagtt ctactaacta acgagtaata tttaaatttt cagcatctcg
     6961 cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc
     7021 cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt
     7081 tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa
     7141 ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc
     7201 caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt
     7261 tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta
     7321 tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat
     7381 cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta
     7441 attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc
     7501 gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg
     7561 tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg
     7621 tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt
     7681 tcctactagt cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7741 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7801 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     7861 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     7921 atgcgtaagg agaaaatacc gcatcaggcg gccttaatta agggcctcgt gatacgccta
     7981 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     8041 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     8101 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     8161 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     8221 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     8281 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     8341 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     8401 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     8461 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     8521 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     8581 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     8641 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     8701 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     8761 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     8821 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     8881 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     8941 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     9001 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     9061 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     9121 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     9181 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     9241 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     9301 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     9361 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     9421 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     9481 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     9541 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     9601 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     9661 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     9721 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     9781 agc
//
