LOCUS       sequence-332384-        9341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of YFP-CaCyclOp(A-2x) in BWMs of C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9341)
  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 9341)
  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: pTH32 [pmyo-3::YFP-CaCyclOp(A-2x)::SL2::mCherry]
FEATURES             Location/Qualifiers
     source          1..9341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2419..2421,2422..2424,2425..3135)
                     /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(2464..2485)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2725..2744)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3072..3093
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5294..6004
                     /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(5439..5457)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5707..5726
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6920..6942)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7042..7061
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7143..7161)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7229..7333
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7334..7402,7403..8194)
                     /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(7552..7571)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8365..8953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8854..8873
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9107..9124
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9241..9262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9277..9282,9283..9300,9301..9307)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9315..9331
                     /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     join(9320..9341,1)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(9339..9341,1..14)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(9339..9341,1..14)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaaacagcta tgaccatgat tacgccaagc ttgcggctat aataagttct tgaataaaat
       61 aattttcccg acaaaacatg agtatttctt tcgaaaataa aagtgcaggc taattagaga
      121 ttattctgta attaactgca taatttgtca cgtgccatag ttttacattc cactacgtca
      181 tagttcttaa aatactaatc tcctgaaaat agaagtaggt gaagaaagtt taattatcag
      241 ttctaaaatg acaattgatc tttggaatat gttctgaaac taccgatcat tgaacagatg
      301 ctatttgaat gatatagaat tgtatatttg caatttctga aacgcgttct taaaggcaca
      361 cagattaatt caaaagggtc tggccgcaaa aaggtttatg gtggccgatt ttgagttttg
      421 tgtgtgattg ctttttcaca atcagtgttt tcaggattat gtgatgaact agatcttcaa
      481 gtttcgttac atttcatatg ttttcggaac tcacgaagta catattgggt attgtgctca
      541 aaaaattcag caatcagctt cgctccgctg actttagaac ccaaaaaaat agtatggcca
      601 aactgactgt gttacgatca tttcaatttt tcaatacata tttaagattt ctaagagtaa
      661 gaaggtcaaa aactgttctg gaatacatat atatttttca ggttacaatt agtcaaaaag
      721 tgcactgaaa tatacgtttt aatttcacga ataacccaat tagttcaatg tatttttggt
      781 caaccaacgt taaagtttgg cttccaacca attatcattt ctgatcaacc acaatgtttt
      841 ttctttatct gcaagttaat tttttatttt tatccagatg tttggcatat ttttcaattc
      901 ttcactagcg cccacttctt gcacttccgg cgccctgaat ctaatgcatc tgttgcaaga
      961 attgaaagac caatcaacac attgttttct tcacgagata ctgaagaaaa tgaataaaaa
     1021 cagagaaaaa gagccatgtg attagtgaca actgttgcta acagatacca tagcttggac
     1081 ttggtacgtg atggcaacgt atgggtcaac aaaaatgatt gcagaggggg tgcaaaacag
     1141 tcaagtcgag aaaatatgaa aaacagaaaa caaagaacag aaaaatgggt ttgagagtca
     1201 gtataattta taaaagaaaa attgtacata gaaattaacc atttttgtag aagaagttat
     1261 ttttcaagca tcgttaaaaa ttattcaaag caccttattt catatttaat tttaaacatg
     1321 gttaaatgaa caacacggtg cgcaatcagg aaaacttgaa atctgaaact gttgttgtga
     1381 tcttcttcgc aactgttcag atagcactag tgtaatgtta agagtgcgcg aatataatgg
     1441 aatataatgg atcacacctc ctgccatcag gtaaacgtct ctgttatcac atatttccaa
     1501 ctattaaatt tttacctttt acagttttac atttttttga aaaaagtaac tttttgtctt
     1561 caaaatccct gacgaaaata tcaaatattt taatcgagac tgcagaggaa ccgattgatg
     1621 atttggaaaa tccagcttta cctgtgtaag aactgaaaag tttcataacc ctagggtatt
     1681 cccagttaca ttccccactg gctaacaata gcacccagtt tttcatcacc tttcttcaaa
     1741 tttctcggcg atttgttaaa aacaaaattt gtgtcccttc tctgatatct ctatgtctct
     1801 aaacacaagt tcatcggaaa acgaaggagg gtaggtgttg gttgggctcc cgaagtgaaa
     1861 atagaagagc aagaatagaa tattagagag agagtgcaga gagggcggga tagctcccgg
     1921 gattccgttt tcttcttctt tatcttcaac gatgatgtgt gtgcgtgttg tatagattct
     1981 gttgctcccc cacaactcgc tccgaaggct caatacaatt caattgatat tggaggagag
     2041 cctaccggag tgggaggata agaagaaaca taagaagaag aagaagaaga agcatgcttc
     2101 tggtttttga tgctatgaaa acggcacaaa aagatgattg aggtcccttt tcaatacctt
     2161 ctctcatctt tcaaatccca ttgaaaccta aaacttctca ccacgcttta ccattgttct
     2221 ccaaaaactt atagcaatgt ctataacttt tttatctctg aaaagcagtg ttccattttt
     2281 ctttttccta ttttatttca attgtttctc acatttcgtt tggattcttt gcttgtcaac
     2341 cagcttcttc ttccactttt accgtctaat tttcagggca gggagccatc aaacccacga
     2401 ccactagatc catctagaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2461 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2521 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
     2581 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc
     2641 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2701 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2761 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2821 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
     2881 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     2941 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3001 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     3061 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3121 gacgagctgt acaagggatc aatgtctatg aaagataaag ataataacct gcggggagca
     3181 tgtagtggct gttcctgccc tgaatactgt tactcaccaa cctcaaccct gtgcgacgat
     3241 tgcaagtgta gcgtgacaaa acaccctatc gtcgagcagc cactgactcg aaacggaagt
     3301 ttccggagct ccggagcctc actgctgcca tctcccagtc agcccaatat caaggtcact
     3361 ggctctagta ccgcctcaag caacgctaat atgcggaaca gacagaacaa ttcactgagc
     3421 gtgtccaatg tccgcagcac ctcctctgca agttcaagca acgtgtcctc tcccgccaat
     3481 agccgacctg gctctcctag taagcagtcc gctctgcagc agtaccagac aaacatcgca
     3541 gacatgtggt cctgggatat gatgctgagt accccttcac tgaaattcct gacaggacag
     3601 tttatcatgt gggccattct gaccgtggcc ggcgctttct acgctctgtt tatccaggaa
     3661 agacaggctt ataatagggg ctgggcagac atttggtacg gctatggggc cttcggattt
     3721 ggcatcggga ttgctttcag ttatatggga tttgcaggcg ccagaaaccc agagaagaaa
     3781 gcactgtccc tgtgcctgct gggagtgaac atcatcgctt ttagttcata catcctgatt
     3841 atgctgcggc tgactccaac catcgaaggc accctgagca acccagtgga gcctgcaaga
     3901 tacctggagt ggattgctac ctgtcccgtc ctgattctgc tgatctccga gattacacag
     3961 gcagaccata atgcctgggg ggtggtcttc tctgattacg ccctggtggt ctgcgggttc
     4021 tttggagctg tgctgccccc ttatccctgg ggaaacctgt ttaatatcct gtcatgtgcc
     4081 ttctttagct tcgtggtcta cagcctgtgg aggtccttta ctggggccat caacggagag
     4141 accccttgca acattgaagt gaatggcctg aggtggacac gcttctcaac tgtcaccaca
     4201 tggaccctgt ttccactgag ctggttcgcc tttacatctg gcatgctgag tttcacaatg
     4261 actgaggcat cttttaccat gatcgatatt ggggccaagg tgttcctgac actggtgctg
     4321 gtcaacagca ctgtggagca ggcacagaat cagaaagtcg aagccatcac agctattgca
     4381 gaggaactgg aatctcagat cactaactgt gacgccattc tgcagaagat gatgcctgag
     4441 ggggtgctgg aacagctgaa gaatggacag gcaaccgagg ccaaagagta cgaatccgtg
     4501 acagtcttct tttctgacat cacaaacttt actgtgatta gctccaggac ctccacaaag
     4561 gacatgatgg ccaccctgaa caagctgtgg ctggagtatg atgccatcgc taagcggtgg
     4621 ggcgtgtata aggtcaagac tattggggac gcttatctgg gagtgaccgg agcaccagag
     4681 gtggtccccg accacgctga tcgagcagtg aacttcgccc tggacatcat cgagatgatc
     4741 aagactttta aaactgctac cggcgaatct atcaacattc gcatcggcct gaatagtgga
     4801 ccagtgacag ccggagtcct gggcgacctg aaccctcatt gggatctggt gggggatacc
     4861 gtcaatacag cttcccggat ggagtcaacc agcaaggcag ggcacattca tatctccgac
     4921 tctacttacc agatgatcaa gggaaaattc gtgacccagc cactggacct gatggaagtg
     4981 aaaggaaagg gaaaaatgca gacatattgg gtgaccgcta gaaagtaagg taccgctgtc
     5041 tcatcctact ttcacctagt taactgcttg tcttaaaatc tatgcttctc tttagtatct
     5101 aaaattttcc tagaagctta caagtatata aatggtctct tctcaataaa ggttgtatat
     5161 ttattcatct tattgaatct gccatttcct cgtttttgcg agtttatata ccttccaatt
     5221 ttctttctat tgtattttca acttctaatt ttaattcagg gaaactgctg taccggtaga
     5281 aaaaatgctc gagatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     5341 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     5401 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5461 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     5521 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     5581 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     5641 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     5701 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     5761 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     5821 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     5881 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     5941 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     6001 gtaagaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
     6061 ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat
     6121 ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca
     6181 tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact
     6241 tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta
     6301 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     6361 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     6421 tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     6481 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     6541 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     6601 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg
     6661 tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     6721 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     6781 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     6841 tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     6901 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6961 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     7021 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     7081 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaattaa
     7141 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     7201 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7261 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7321 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7381 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7441 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7501 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7561 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7621 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7681 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7741 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7801 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7861 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7921 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7981 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8041 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8101 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8161 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8221 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8281 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8401 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8461 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8521 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8581 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8641 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8701 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8761 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8821 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8881 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8941 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9001 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9061 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     9121 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     9181 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     9241 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     9301 tatgttgtgt ggaattgtga gcggataaca atttcacaca g
//
