LOCUS       sequence-332387-        9930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of YFP-BeCyclOp(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 9930)
  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 9930)
  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: pTH41 [punc-17::YFP-BeCyclOp(A-2x)::SL2::mCherry]
FEATURES             Location/Qualifiers
     source          1..9930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(3213..3215,3216..3218,3219..3929)
                     /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(3258..3279)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3519..3538)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3866..3887
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6235..6945
                     /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(6380..6398)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6648..6667
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7861..7883)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7983..8002
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8084..8102)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8170..8274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8275..8343,8344..9135)
                     /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(8493..8512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9306..9894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9795..9814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgggatta caccaatcat ttctcccctc cgaaatgtgt
       61 agtcgtcatg acaaagtggt gacactggat tacggtaggc tccgcccact tcttggaaaa
      121 gtgacgtggg ctcggtgagc cctaaagctt ttggttttca caattttctg gtttttggga
      181 tttttgcggg aattttgaga ggagaaagtt gggagagaag atattggaat ggtccccggg
      241 gagctgtaca tttatacatc tagactgaag ctggggtatt gcaaaaagag aaaagcctat
      301 cactttcttt ccatcctgca tacttgccaa aaagttttgg ctgagcttat ttttaaaatt
      361 ggaatttttt ggaattttca gaaaagaaaa agttttttct ttttttttaa tttaaaaaat
      421 ttttgctatt tattgcatgg tactacagaa aaacttgaat gaaataaaaa atttggaaaa
      481 ttcaagaaaa cttttttttt catagtttat ttattcgagt tttaatgtgg cttttatttt
      541 gttcggtttt tgttttaaaa agattttata caaaatttca agaatttaca atttttccgt
      601 tgtaaaaaat ccaaaaaaaa atccaaaaaa atagttcaga ccttttttta aattttaaat
      661 ttaaaaattc aataccaaaa aattaaaaaa aattagtttt tccgagaata aaggaaaaat
      721 gttgaaaaaa tattatatat aaaatttcca gaaaatgtga aaactccaaa aaactgcaga
      781 cttttcccca aactagctat tttttgaatt cattaccttt aaaggtggag taccgaaatt
      841 taagactttg cttttttaga cccaaaatgg tccaaaacta ccaaatttcg taatgagacg
      901 ttctgaaaat ttttcaaaaa aatgttatgg gagctcaaag tttttgaaaa aatggtatat
      961 ttttagctaa ttttaaccac ttcttggcgt cgcagcggtt ggaactcaat tttaattaaa
     1021 ttatcatcct tcaatgtcct tcaatctttg caatttcaat cgaccatacg cgttcagagc
     1081 attttttttt ttcgtttttc ctcccgtcca gcacatattt aaatattctc tctccccctc
     1141 ccctggttcc taaaaacttg ggaaagggga atttcgaagg aggtggaagc ttcttcttct
     1201 ccccgagcac acacacacat cctctcttca aaatctttaa gaaatagtgc gtgtgaatga
     1261 ctagacgctg aagctaactt ggtttccgcg aacaatccca atcagtgcct gtgccgaggc
     1321 ttcggagaag aagagattga cgtcgtcttt ttcttttttt gtgaaacaga ataaaaaaga
     1381 atacgaaatg gactttttca cacattttcg ttacttttcc agaaaaaaac gaagctctag
     1441 ccatggaagt ttgaattgaa aacctaaacc ccgatgtgta ttcatttgag cttaagccta
     1501 agcctaattc tacgcctaag cataagccta agcctatgtt caagcctaag ccaaattgta
     1561 gaacgttacc aaaagatcaa acctattttt cctacagaaa aggtctaatg ataactgtta
     1621 agcgagagac aaggagagat atatatcaaa aactagccca aactgtgatt aacatctatt
     1681 aactgacctc aaaaaaaaaa atagtagaga ttaaatactg ctcaaaaagt ttcttggaaa
     1741 agtaaattgt taggtttctg gggcctgtaa aactggaccg agttttgaag aattttttta
     1801 acttttcaaa gtttggcgtc actgaaagct ttatattgta catgttccac caattttctt
     1861 ttgtatccgt tttttctgtc tcaaaacgga gctcgagccc atgacgtgtg gataccgtga
     1921 gaatccgccc aattccttca catcgatctc atatttcttt cccagtttat attgaattcc
     1981 cgttgaattg cggctttgca ggggagaaga tgtgcgtctt tatatttttt tttctatcct
     2041 tacccgccgt gaatccggct cgaattgagc caatttttat gaatagtaga gctaattgac
     2101 ttcgtctccc tagaattgat gaggtaaagc tcgtggggag atatggagat ttttcgattt
     2161 tttttccagt aattttcagt tcaaaatttg agcttttgag caggagtttt gagatttgtg
     2221 ccaaaatttc ctattatttt ttcaaatatt tttgcttttg taaaaaaggt gtagtagaag
     2281 ttttgtagtg ccactatttt tttttgaaaa ttatgttttg gcggaaagtt ctttaaaaaa
     2341 gttaatcaaa atttttggcg ctttagtcaa attaattttc tggcaaaaat attagtaatt
     2401 ttggcgtgag ttcaattttc ccattcttaa aatttcacac aattaagaat tttaagattt
     2461 gggttttttt ttcaacaaaa aatcaaatat ttacatgaaa attcggcggg aaataaaaaa
     2521 aaatattccg attcaatgtg tgaataactt tttgcgggga attcaaattt gcattattta
     2581 aacttttttt aagctcgaaa aattgaaatt cttaaataaa taaataaaat tttcctcgaa
     2641 ttttgaatgt ttcgattttt tagcgttatt ttttcaaaat ttccactaaa actctagttc
     2701 taaagttcta aaactttaaa aacttcctaa tgttgtggtc ttgtgttttt ttggcagcga
     2761 tttccaaatt tctgaaattc caacaaaaaa accactacgt cagtttgcat atccaacccc
     2821 cccccccccc ccctctgagc caatcaatca tgtgcatttt gaattggcgc caaggcaatc
     2881 ttcgtaaaaa ataacaatag agtcatccct cagaacagaa aagaaacccg cccgggggct
     2941 caaatacaca tgtcattttt cttttttcgc ctgattttta gcctaaatta gtgtttagca
     3001 attttttaag gtaactcggt ggagtcattt gaacttttta aactaaaatt tcagaaaaaa
     3061 ttcttttttt ttgtcaaaaa ataattttcc cgccggaatt tttcaaaaaa ataattttta
     3121 ttttattttt caagttcctt gtttcctctt tattttttta ttttgccccc aaaagtttta
     3181 cactaaaata tttaattttc agctcggcta gcatggtgag caagggcgag gagctgttca
     3241 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3301 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     3361 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc
     3421 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3481 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3541 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3601 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3661 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     3721 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3781 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca
     3841 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3901 tcactctcgg catggacgag ctgtacaagg gatcaatgaa ggacaaggac aacaacctcc
     3961 gtggagcctg ctcctcctgc aactgcccag agtactgctt ctccccaacc tccaccctct
     4021 gcgacgactg caagtgctcc gtcaccaagc acccaatcgt cgagcaacca ctctcccgta
     4081 acggatcctt ccgttcctcc ggagcctccc tcctcccatc cccatcctcc ccaaacgtca
     4141 agatcacctc caccgtcgga ctccgttccc gtaagtccga gtcccaagcc aacgtccgtg
     4201 gatccatgat ctccaactcc aactccggat cccgttccaa caactccgga ggagccggag
     4261 gaggatccgg aggatcctcc tcctccaagg taagtttaaa catatatata ctaactaacc
     4321 ctgattattt aaattttcag ggaggatccg ccctcgccaa ctaccaatcc gccatgtccg
     4381 agctctggtc ctggaacatg atgctctcca ccccatccct caagttcctc accgtccaat
     4441 tcaccacctg gatcgtcctc accaccgtcg gagccatcta caccctcttc ttccacgagc
     4501 gccaagccta caaccgtgga tgggccgaca tctggtacgg atacggagcc ttcggattcg
     4561 gactcggact ctccttcgcc tacatgggat tcaccggagc ccgtaaccca gagaagaagg
     4621 ccctctccct ctgcctcctc ggagtcaact tcatctcctt catgtcctac atcatcatca
     4681 tgctccgtct caccccaacc atcgagggaa ccatggccaa cccagtcgag ccagcccgtt
     4741 acctcgagtg gatcgccacc tgcccagtcc ttatcctcct catctccgag atcacccaat
     4801 acccacacga cccatacaag gtaagtttaa acagttcggt actaactaac catacatatt
     4861 taaattttca ggtcatcgtc aacgactacg ccctctgcct cgccggattc gtcggagcca
     4921 tctccgccca acaaccatgg ggagacctcg cccacttcgt ctcctgcctc tgcttctcct
     4981 acgtcgtcta ctccctctgg tcctgcttca ccggagccat cgacggagag acccaatgca
     5041 acgtcgagaa gtccggactc cgttggatcc gtttctccac catcaccacc tggtccctct
     5101 tcccaatcac ctggttctcc tacacctccg gactcatctc cttcaccgtc gccgaggccg
     5161 gattctccat gatcgacatc ggagccaagg tcttcctcac cctcgtcctc gtcaactcca
     5221 ccgtcgagca agcccaaaac caaaaggtcg acgccatcac cgccatcgcc gaggagctcg
     5281 agaaccaaat caacaactgc gacgccatcc tccaaaagat gatgccagag ggagtcctcg
     5341 agcaactcaa gaacggacaa gccaccgagg ccaaggagta cgagtccgtc accgtcttct
     5401 tctccgacat caccaacttc accgtcatct cctcccgtac ctccaccaag gtaagtttaa
     5461 acatgatttt actaactaac taatctgatt taaattttca ggacatgatg gccaccctca
     5521 acaagctctg gctcgagtac gacgccatcg ccaagcgttg gggagtctac aaggtcaaaa
     5581 ccatcggaga cgcctacctc ggagtcaccg gagccccaga cgtcgtccca gaccacgccg
     5641 agcgtgcctg caacttcgcc gtcgacatca tcgagatgat caagtccttc aagaccatca
     5701 ccggagagtc catcaacatc cgtatcggac tcaactccgg accagtcacc gccggagtcc
     5761 tcggagacct caacccacac tgggacctcg tcggagacac cgtcaacacc gcctcccgta
     5821 tggagtccac ctccaaggcc ggacacatcc acatctccga gtccacctac cacttcatca
     5881 agtccaagtt cgtcacccaa ccactcgacg tcatggaggt caagggaaag ggaaagatgc
     5941 aaacctactg ggtcctcgga cgtaagtaag gtaccgctgt ctcatcctac tttcacctag
     6001 ttaactgctt gtcttaaaat ctatgcttct ctttagtatc taaaattttc ctagaagctt
     6061 acaagtatat aaatggtctc ttctcaataa aggttgtata tttattcatc ttattgaatc
     6121 tgccatttcc tcgtttttgc gagtttatat accttccaat tttctttcta ttgtattttc
     6181 aacttctaat tttaattcag ggaaactgct gtaccggtag aaaaaatgct cgagatggtg
     6241 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     6301 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     6361 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     6421 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     6481 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     6541 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     6601 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     6661 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     6721 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     6781 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     6841 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     6901 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagaatt ccaactgagc
     6961 gccggtcgct accattacca acttgtctgg tgtcaaaaat aataggggcc gctgtcatca
     7021 gagtaagttt aaactgagtt ctactaacta acgagtaata tttaaatttt cagcatctcg
     7081 cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc
     7141 cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt
     7201 tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa
     7261 ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc
     7321 caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt
     7381 tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta
     7441 tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat
     7501 cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta
     7561 attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc
     7621 gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg
     7681 tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg
     7741 tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt
     7801 tcctactagt cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7861 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7921 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     7981 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     8041 atgcgtaagg agaaaatacc gcatcaggcg gccttaatta agggcctcgt gatacgccta
     8101 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     8161 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     8221 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     8281 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     8341 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     8401 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     8461 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     8521 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     8581 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     8641 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     8701 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     8761 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     8821 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     8881 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     8941 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     9001 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     9061 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     9121 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     9181 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     9241 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     9301 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     9361 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     9421 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     9481 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     9541 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     9601 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     9661 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     9721 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     9781 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     9841 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     9901 agcaacgcgg cctttttacg gttcctggcc
//
