LOCUS       sequence-332385-        9488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of YFP-BeCyclOp(A-2x) in BWMs of C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9488)
  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 9488)
  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: pTH33 [pmyo-3::YFP-BeCyclOp(A-2x)::SL2::mCherry]
FEATURES             Location/Qualifiers
     source          1..9488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    13..34
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(49..54,55..72,73..79)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    87..103
                     /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     92..114
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     111..127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     111..127
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(2532..2534,2535..2537,2538..3248)
                     /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(2577..2598)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2838..2857)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3185..3206
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5554..6264
                     /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(5699..5717)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5967..5986
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7180..7202)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7302..7321
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7403..7421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7489..7593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7594..7662,7663..8454)
                     /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(7812..7831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8625..9213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9114..9133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9367..9384
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
       61 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      121 ctatgaccat gattacgcca agcttgcggc tataataagt tcttgaataa aataattttc
      181 ccgacaaaac atgagtattt ctttcgaaaa taaaagtgca ggctaattag agattattct
      241 gtaattaact gcataatttg tcacgtgcca tagttttaca ttccactacg tcatagttct
      301 taaaatacta atctcctgaa aatagaagta ggtgaagaaa gtttaattat cagttctaaa
      361 atgacaattg atctttggaa tatgttctga aactaccgat cattgaacag atgctatttg
      421 aatgatatag aattgtatat ttgcaatttc tgaaacgcgt tcttaaaggc acacagatta
      481 attcaaaagg gtctggccgc aaaaaggttt atggtggccg attttgagtt ttgtgtgtga
      541 ttgctttttc acaatcagtg ttttcaggat tatgtgatga actagatctt caagtttcgt
      601 tacatttcat atgttttcgg aactcacgaa gtacatattg ggtattgtgc tcaaaaaatt
      661 cagcaatcag cttcgctccg ctgactttag aacccaaaaa aatagtatgg ccaaactgac
      721 tgtgttacga tcatttcaat ttttcaatac atatttaaga tttctaagag taagaaggtc
      781 aaaaactgtt ctggaataca tatatatttt tcaggttaca attagtcaaa aagtgcactg
      841 aaatatacgt tttaatttca cgaataaccc aattagttca atgtattttt ggtcaaccaa
      901 cgttaaagtt tggcttccaa ccaattatca tttctgatca accacaatgt tttttcttta
      961 tctgcaagtt aattttttat ttttatccag atgtttggca tatttttcaa ttcttcacta
     1021 gcgcccactt cttgcacttc cggcgccctg aatctaatgc atctgttgca agaattgaaa
     1081 gaccaatcaa cacattgttt tcttcacgag atactgaaga aaatgaataa aaacagagaa
     1141 aaagagccat gtgattagtg acaactgttg ctaacagata ccatagcttg gacttggtac
     1201 gtgatggcaa cgtatgggtc aacaaaaatg attgcagagg gggtgcaaaa cagtcaagtc
     1261 gagaaaatat gaaaaacaga aaacaaagaa cagaaaaatg ggtttgagag tcagtataat
     1321 ttataaaaga aaaattgtac atagaaatta accatttttg tagaagaagt tatttttcaa
     1381 gcatcgttaa aaattattca aagcacctta tttcatattt aattttaaac atggttaaat
     1441 gaacaacacg gtgcgcaatc aggaaaactt gaaatctgaa actgttgttg tgatcttctt
     1501 cgcaactgtt cagatagcac tagtgtaatg ttaagagtgc gcgaatataa tggaatataa
     1561 tggatcacac ctcctgccat caggtaaacg tctctgttat cacatatttc caactattaa
     1621 atttttacct tttacagttt tacatttttt tgaaaaaagt aactttttgt cttcaaaatc
     1681 cctgacgaaa atatcaaata ttttaatcga gactgcagag gaaccgattg atgatttgga
     1741 aaatccagct ttacctgtgt aagaactgaa aagtttcata accctagggt attcccagtt
     1801 acattcccca ctggctaaca atagcaccca gtttttcatc acctttcttc aaatttctcg
     1861 gcgatttgtt aaaaacaaaa tttgtgtccc ttctctgata tctctatgtc tctaaacaca
     1921 agttcatcgg aaaacgaagg agggtaggtg ttggttgggc tcccgaagtg aaaatagaag
     1981 agcaagaata gaatattaga gagagagtgc agagagggcg ggatagctcc cgggattccg
     2041 ttttcttctt ctttatcttc aacgatgatg tgtgtgcgtg ttgtatagat tctgttgctc
     2101 ccccacaact cgctccgaag gctcaataca attcaattga tattggagga gagcctaccg
     2161 gagtgggagg ataagaagaa acataagaag aagaagaaga agaagcatgc ttctggtttt
     2221 tgatgctatg aaaacggcac aaaaagatga ttgaggtccc ttttcaatac cttctctcat
     2281 ctttcaaatc ccattgaaac ctaaaacttc tcaccacgct ttaccattgt tctccaaaaa
     2341 cttatagcaa tgtctataac ttttttatct ctgaaaagca gtgttccatt tttctttttc
     2401 ctattttatt tcaattgttt ctcacatttc gtttggattc tttgcttgtc aaccagcttc
     2461 ttcttccact tttaccgtct aattttcagg gcagggagcc atcaaaccca cgaccactag
     2521 atccatctag aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2581 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2641 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     2701 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg
     2761 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2821 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2881 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2941 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     3001 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     3061 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3121 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     3181 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     3241 tgtacaaggg atcaatgaag gacaaggaca acaacctccg tggagcctgc tcctcctgca
     3301 actgcccaga gtactgcttc tccccaacct ccaccctctg cgacgactgc aagtgctccg
     3361 tcaccaagca cccaatcgtc gagcaaccac tctcccgtaa cggatccttc cgttcctccg
     3421 gagcctccct cctcccatcc ccatcctccc caaacgtcaa gatcacctcc accgtcggac
     3481 tccgttcccg taagtccgag tcccaagcca acgtccgtgg atccatgatc tccaactcca
     3541 actccggatc ccgttccaac aactccggag gagccggagg aggatccgga ggatcctcct
     3601 cctccaaggt aagtttaaac atatatatac taactaaccc tgattattta aattttcagg
     3661 gaggatccgc cctcgccaac taccaatccg ccatgtccga gctctggtcc tggaacatga
     3721 tgctctccac cccatccctc aagttcctca ccgtccaatt caccacctgg atcgtcctca
     3781 ccaccgtcgg agccatctac accctcttct tccacgagcg ccaagcctac aaccgtggat
     3841 gggccgacat ctggtacgga tacggagcct tcggattcgg actcggactc tccttcgcct
     3901 acatgggatt caccggagcc cgtaacccag agaagaaggc cctctccctc tgcctcctcg
     3961 gagtcaactt catctccttc atgtcctaca tcatcatcat gctccgtctc accccaacca
     4021 tcgagggaac catggccaac ccagtcgagc cagcccgtta cctcgagtgg atcgccacct
     4081 gcccagtcct tatcctcctc atctccgaga tcacccaata cccacacgac ccatacaagg
     4141 taagtttaaa cagttcggta ctaactaacc atacatattt aaattttcag gtcatcgtca
     4201 acgactacgc cctctgcctc gccggattcg tcggagccat ctccgcccaa caaccatggg
     4261 gagacctcgc ccacttcgtc tcctgcctct gcttctccta cgtcgtctac tccctctggt
     4321 cctgcttcac cggagccatc gacggagaga cccaatgcaa cgtcgagaag tccggactcc
     4381 gttggatccg tttctccacc atcaccacct ggtccctctt cccaatcacc tggttctcct
     4441 acacctccgg actcatctcc ttcaccgtcg ccgaggccgg attctccatg atcgacatcg
     4501 gagccaaggt cttcctcacc ctcgtcctcg tcaactccac cgtcgagcaa gcccaaaacc
     4561 aaaaggtcga cgccatcacc gccatcgccg aggagctcga gaaccaaatc aacaactgcg
     4621 acgccatcct ccaaaagatg atgccagagg gagtcctcga gcaactcaag aacggacaag
     4681 ccaccgaggc caaggagtac gagtccgtca ccgtcttctt ctccgacatc accaacttca
     4741 ccgtcatctc ctcccgtacc tccaccaagg taagtttaaa catgatttta ctaactaact
     4801 aatctgattt aaattttcag gacatgatgg ccaccctcaa caagctctgg ctcgagtacg
     4861 acgccatcgc caagcgttgg ggagtctaca aggtcaaaac catcggagac gcctacctcg
     4921 gagtcaccgg agccccagac gtcgtcccag accacgccga gcgtgcctgc aacttcgccg
     4981 tcgacatcat cgagatgatc aagtccttca agaccatcac cggagagtcc atcaacatcc
     5041 gtatcggact caactccgga ccagtcaccg ccggagtcct cggagacctc aacccacact
     5101 gggacctcgt cggagacacc gtcaacaccg cctcccgtat ggagtccacc tccaaggccg
     5161 gacacatcca catctccgag tccacctacc acttcatcaa gtccaagttc gtcacccaac
     5221 cactcgacgt catggaggtc aagggaaagg gaaagatgca aacctactgg gtcctcggac
     5281 gtaagtaagg taccgctgtc tcatcctact ttcacctagt taactgcttg tcttaaaatc
     5341 tatgcttctc tttagtatct aaaattttcc tagaagctta caagtatata aatggtctct
     5401 tctcaataaa ggttgtatat ttattcatct tattgaatct gccatttcct cgtttttgcg
     5461 agtttatata ccttccaatt ttctttctat tgtattttca acttctaatt ttaattcagg
     5521 gaaactgctg taccggtaga aaaaatgctc gagatggtga gcaagggcga ggaggataac
     5581 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     5641 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     5701 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     5761 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     5821 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     5881 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     5941 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     6001 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     6061 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     6121 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     6181 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     6241 ggcatggacg agctgtacaa gtaagaattc caactgagcg ccggtcgcta ccattaccaa
     6301 cttgtctggt gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc
     6361 tactaactaa cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta
     6421 agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt
     6481 tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc
     6541 tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa
     6601 aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca caatgttctg
     6661 tgtacacttc ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc
     6721 gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt
     6781 cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat
     6841 ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt
     6901 ggggtttccc ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca
     6961 agtattgatg agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg
     7021 aaggtgagta gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccttaa
     7081 atgacagaat acattcccaa tataccaaac ataactgttt cctactagtc ggccgtacgg
     7141 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7201 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7261 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7321 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7381 catcaggcgg ccttaattaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     7441 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     7861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     8041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     8101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     8161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     8221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     8341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8761 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     8821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     9001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     9061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     9121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     9181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     9241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     9301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     9481 ggcagtga
//
