LOCUS       sequence-332383-        8804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of BeCNG1-YFP in BWMs of C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8804)
  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 8804)
  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: pASH3 [pmyo-3::BeCNG1-YFP]
FEATURES             Location/Qualifiers
     source          1..8804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2440..2449
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4633..4642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4639..4641,4642..4644,4645..5358)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4684..4705)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4945..4964)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5292..5313
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6275..6297)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6397..6416
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6494..6512)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6580..6684
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6685..6753,6754..7545)
                     /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(6903..6922)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7716..8304
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8205..8224
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8458..8475
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8592..8613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8628..8633,8634..8651,8652..8658)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8666..8682
                     /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     8671..8693
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8690..8706
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8690..8706
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gatggatacg ctaacaactt ggaaatgaaa taagcttggg ctgcaggtcg gctataataa
       61 gttcttgaat aaaataattt tcccgacaaa acatgagtat ttctttcgaa aataaaagtg
      121 caggctaatt agagattatt ctgtaattaa ctgcataatt tgtcacgtgc catagtttta
      181 cattccacta cgtcatagtt cttaaaatac taatctcctg aaaatagaag taggtgaaga
      241 aagtttaatt atcagttcta aaatgacaat tgatctttgg aatatgttct gaaactaccg
      301 atcattgaac agatgctatt tgaatgatat agaattgtat atttgcaatt tctgaaacgc
      361 gttcttaaag gcacacagat taattcaaaa gggtctggcc gcaaaaaggt ttatggtggc
      421 cgattttgag ttttgtgtgt gattgctttt tcacaatcag tgttttcagg attatgtgat
      481 gaactagatc ttcaagtttc gttacatttc atatgttttc ggaactcacg aagtacatat
      541 tgggtattgt gctcaaaaaa ttcagcaatc agcttcgctc cgctgacttt agaacccaaa
      601 aaaatagtat ggccaaactg actgtgttac gatcatttca atttttcaat acatatttaa
      661 gatttctaag agtaagaagg tcaaaaactg ttctggaata catatatatt tttcaggtta
      721 caattagtca aaaagtgcac tgaaatatac gttttaattt cacgaataac ccaattagtt
      781 caatgtattt ttggtcaacc aacgttaaag tttggcttcc aaccaattat catttctgat
      841 caaccacaat gttttttctt tatctgcaag ttaatttttt atttttatcc agatgtttgg
      901 catatttttc aattcttcac tagcgcccac ttcttgcact tccggcgccc tgaatctaat
      961 gcatctgttg caagaattga aagaccaatc aacacattgt tttcttcacg agatactgaa
     1021 gaaaatgaat aaaaacagag aaaaagagcc atgtgattag tgacaactgt tgctaacaga
     1081 taccatagct tggacttggt acgtgatggc aacgtatggg tcaacaaaaa tgattgcaga
     1141 gggggtgcaa aacagtcaag tcgagaaaat atgaaaaaca gaaaacaaag aacagaaaaa
     1201 tgggtttgag agtcagtata atttataaaa gaaaaattgt acatagaaat taaccatttt
     1261 tgtagaagaa gttatttttc aagcatcgtt aaaaattatt caaagcacct tatttcatat
     1321 ttaattttaa acatggttaa atgaacaaca cggtgcgcaa tcaggaaaac ttgaaatctg
     1381 aaactgttgt tgtgatcttc ttcgcaactg ttcagatagc actagtgtaa tgttaagagt
     1441 gcgcgaatat aatggaatat aatggatcac acctcctgcc atcaggtaaa cgtctctgtt
     1501 atcacatatt tccaactatt aaatttttac cttttacagt tttacatttt tttgaaaaaa
     1561 gtaacttttt gtcttcaaaa tccctgacga aaatatcaaa tattttaatc gagactgcag
     1621 aggaaccgat tgatgatttg gaaaatccag ctttacctgt gtaagaactg aaaagtttca
     1681 taaccctagg gtattcccag ttacattccc cactggctaa caatagcacc cagtttttca
     1741 tcacctttct tcaaatttct cggcgatttg ttaaaaacaa aatttgtgtc ccttctctga
     1801 tatctctatg tctctaaaca caagttcatc ggaaaacgaa ggagggtagg tgttggttgg
     1861 gctcccgaag tgaaaataga agagcaagaa tagaatatta gagagagagt gcagagaggg
     1921 cgggatagct cccgggattc cgttttcttc ttctttatct tcaacgatga tgtgtgtgcg
     1981 tgttgtatag attctgttgc tcccccacaa ctcgctccga aggctcaata caattcaatt
     2041 gatattggag gagagcctac cggagtggga ggataagaag aaacataaga agaagaagaa
     2101 gaagaagcat gcttctggtt tttgatgcta tgaaaacggc acaaaaagat gattgaggtc
     2161 ccttttcaat accttctctc atctttcaaa tcccattgaa acctaaaact tctcaccacg
     2221 ctttaccatt gttctccaaa aacttatagc aatgtctata acttttttat ctctgaaaag
     2281 cagtgttcca tttttctttt tcctatttta tttcaattgt ttctcacatt tcgtttggat
     2341 tctttgcttg tcaaccagct tcttcttcca cttttaccgt ctaattttca gggcagggag
     2401 ccatcaaacc cacgaccact agatccatct agaggatccg ccaccatggc tgttgaaaac
     2461 gatgctgagg tgcacaccat ggaaaagtct ggtgctgctg ctaaccacaa ctacatctct
     2521 acctactgct gctctcctag atgcgctaga ctccttgaga ctacaagtgc agctgctgct
     2581 gctgcatact ctaggactcc tcttctctct gaaaccgtgc acttcttcag tgagactgga
     2641 agaaggctcg gattcttcac cagggatggt gatgattctg attctccaga cgacgatgat
     2701 gatccagatg atcctgatga tgatcctgct cctgctcttg gacctacagc tgctggtaga
     2761 agaggtgata gggatgatgc tgatcttctc gctggtgcag ctagacctat ggctgatgct
     2821 agagctttgg ctaggatggg aggatcttct ggttctggat ctacccagtc taccagaaaa
     2881 agaaacggtg ctggtggaag gtcatctagg cttgctgatg gacctgataa cgatctcttc
     2941 attgctggaa accagggtgg tggtggactt ttcggaggac ttacttggca ggatctttgg
     3001 cagaagctcc tcaagttcta cctcattcct ggtgagccat tcctcacctg gtgggattac
     3061 ttcatggttc tcgtggctgt gaccaacgga atcactatcc cttttatcgc tgctttcaga
     3121 tacctcttca ccggatcttt catcgtgcag tacgctctcg atgtgatcta ccttatcgat
     3181 atcttcatca agttccacgt gtcatacctc gatagaggat tctacgtgat cttccctaaa
     3241 gaaatggcta tgcactacat gaggtcttgg gagttctctt tcgatctttt caccaacctc
     3301 cctttcgatt tgatcgctct ctcttggtct aactctgctg tgcctggtga ggtgctctac
     3361 ttcttgtcta tcgtgagact ccacaagttg atgaggacct tcaagctcct ctggtggttc
     3421 cacaaagaag agaagagact caacgctcag gctgtgttcc agatgtacaa gttcaccatc
     3481 tacatcaccc tcatcaccca ctggtgtacc tgcatctggt acgttttggc ttgccctgat
     3541 cagtgcgtta caccttcttg ggctactgat cctaaccaga accctgattt ctctgatagg
     3601 ctctctatgt acgtgtacgc actctactgg accgtgatga ctatgactac taccggatac
     3661 ggtgatgtgc acgctaccaa cgatagagag aggatctggt ctatcatctg catgatcacc
     3721 ggtgtgttcc tctacggata cgtgtcaggt actatcgctt cttcactctc taacctcgat
     3781 tcaagaaggg tggcatacag gcagaaggtt gaggctatca agcagtacat gctcgatcag
     3841 aacatggata aggatatgca gaagagggtt accggttggt acgagtacac ttgggagagg
     3901 aacaagggaa tcgatgttat caacgtgttc aacgatctcc catctaactt cagggctgag
     3961 gttgcactct ctgtgaacga ggatatcatc gagaaggctc ctatcttcag aaacacctct
     4021 ctcggattca gaaggatgct ctcaatcgct atgaggatca acttcttcac cgctgatacc
     4081 tacatcgtgc acagaggtga gatcggaaaa gagatgttct tcgttgttca gggaagagtg
     4141 gatgtgctca accacgatga gtctaagcct atctcttcta tggttgaggg atcttacttc
     4201 ggagagtata tgctcatcct cggaaacgtg tgtgaggctt ctgctatcgc tgtgtgcaac
     4261 tgtgatgtgt acgtgctctc taaggatgat tacgatcagg cttgcgctaa ctaccctgag
     4321 gatgctaaca aggttctcgc tgctaccaag gatgcttacc agaagcaact tgagaagcag
     4381 cagaggtcac agagaaagaa atctaccgat gatttcgatc tcggaactac cagtggactc
     4441 cttggagttg gaggtgctac taaccttacc gttatgcctt ctgtgatcat ggaagagatg
     4501 ggatctacta tgggaatggc ttctccacct cagtctggtg ttcaaactag aagggcttca
     4561 ggtgatgatg ctaggaagtc taggcctact ctcactgtgc ctgaagagga tctcgagaag
     4621 gtacccgcgg ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     4681 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     4741 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4801 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     4861 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4921 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     4981 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     5041 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     5101 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     5161 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     5221 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     5281 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     5341 gacgagctgt acaagtaaag aattccaact gagcgccggt cgctaccatt accaacttgt
     5401 ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa gtttaaactg agttctacta
     5461 actaacgagt aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc
     5521 aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt
     5581 tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag tcacctctaa
     5641 caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt
     5701 gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac
     5761 acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca
     5821 caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac
     5881 gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg
     5941 gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt
     6001 ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat
     6061 tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt
     6121 gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac
     6181 agaatacatt cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct
     6241 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6301 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6361 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6421 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6481 ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     6541 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6601 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6661 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6721 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6781 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6841 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6901 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6961 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     7021 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7081 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7141 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7201 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7261 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7321 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7381 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7441 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7501 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7561 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7621 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7681 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7741 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7801 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7861 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7921 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7981 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8041 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8101 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8161 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8221 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8281 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8341 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8401 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8461 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8521 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8581 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8641 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8701 tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc
     8761 atttaaattt tcagagctta aaaatggctg aaatcactca caac
//
