LOCUS       sequence-410713-        9590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pan-neuronal expression of SNG-1::CRY2(D387A)olig(535) in neurons of
            C. elegans. CRY2(D387A) is photoinactive.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9590)
  AUTHORS   Vettkotter D, Schneider M, Goulden BD, Dill H, Liewald J, Zeiler S, 
            Guldan J, Ates YA, Watanabe S, Gottschalk A
  TITLE     Rapid and reversible optogenetic silencing of synaptic transmission 
            by clustering of synaptic vesicles.
  JOURNAL   Nat Commun. 2022 Dec 19;13(1):7827. doi: 10.1038/s41467-022-35324-z.
  PUBMED    36535932
REFERENCE   2  (bases 1 to 9590)
  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: "Nat 
            Commun."; date: "2022-12-19"; volume: "13(1):7827. doi"; pages: " 
            10.1038/s41467-022-35324-z"
COMMENT     Sequence Label: pDV21
            [pSNG-1::SNG-1::CRY2(D387A)olig(535)::SL...
FEATURES             Location/Qualifiers
     source          1..9590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3510..5003
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLAADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
     CDS             5395..6105
                     /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(5540..5558)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5808..5827
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7021..7043)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7143..7162
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7244..7262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7330..7434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7435..7503,7504..8295)
                     /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(7653..7672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8466..9054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8955..8974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9208..9225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9342..9363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9378..9383,9384..9401,9402..9408)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9416..9432
                     /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     9421..9443
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9440..9456
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9440..9456
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgcatgcctg cagagagcgt gttctgttta gggaaatgca tgccctgtac atcatcttcc
       61 gttgaatttt aattacttct actaaacatt tttttgcttg actttttttg caaagactaa
      121 tattttttct gaataaataa gtctgttgtt tcctgcacta ttctggcata tttccttcct
      181 tttaattagt gaagtagttc cttttttcat ctgatgcttg ttttttcaag atacgtcatg
      241 ttctaactga aaacaaacat tttctttaga ttcaattttt ctttagactc aattttaggt
      301 tgacgtttag gttttaaaaa aatttcattc aggtttgttt caaatgttta aaaactttta
      361 ttttccactg aatgaaaact attaatatta agcagtacac aaaatgaaag gtggattagt
      421 gaagtttttt tttctttttt gaagaaaagg cttcatatat gtttaaattg tcaaaaagta
      481 atatcgaaaa ctgaaagggt ttggatcgtt ttgagttgtt tgcagcagta caatttttta
      541 caattacaca tttaatgcag gcaccataat gttttcaaat tgcttagcag gtcagtattc
      601 tatgcaattg tttataaaaa gttatatttt tcatgattga ttctctgttt ccagctcagt
      661 tactgatcaa ttcaatagac gtaatttaat aacaaatatg ccgcgttttt caatttttca
      721 cataagcgtt cttgttcttg accaaatcat ttctctccga aaccgcaaaa tttcccatat
      781 ctgcgccggt tattgaatat tcattggttt gagctcactt ttcatcgttc catagtcggt
      841 ttcctcattc aattttcatt aataattcag tatgcattca ctaaatttgt ctccacggtg
      901 atttaatgaa caagtcttta atgaatttaa tgctgcctgc ggacaagcta gatttagatg
      961 ttctaatgaa tcatcttttt agaattcata tgttattaat gaaacttcct tttatccctc
     1021 tcttttcatt gcaccagaga ctttcaactt ccggaagaaa gaattcgttg aaaagtgctt
     1081 ccaaatcgac tggcttcttt tactacagga ggaccctttg ctctgctttt cgccgtctgt
     1141 cgtttttttt tacgctttcc cttcatggtc gcactgggag gggagccttc tgcgcaaagt
     1201 tgtgggcatg aagagtggct ccgtccttcg tcttcttttt ccccgctact tcttcatctt
     1261 cgctttcggc gtcgtcttct ccttctcggc gagcagagag tgcctctatc tttcttttct
     1321 tcactggatg tgtatatggc gtttagaagt agaagcattc gtgggacgca ctgaatgtct
     1381 tggaagaaag aaaggagaaa agaagctcaa gattacacgc tttcgttctt ttcgttcttt
     1441 tcatttcatc ttttcttttg gtccttccgt ttcaatattt tgtatttaca atgttcttca
     1501 tttcattcca gctcattcca tttaatactt agtatgtcta acagaatctg aacacctctg
     1561 accccagtta tcctaatctc tattgtgtgc aaatctgcat atatttttaa tcctctatat
     1621 ttcttttatt ttagcatgga gaacgtgcgt gcttatggtg ccggactagc tggcgccaat
     1681 ttcgacaaga acacattttt caaaaaacca acggttttgt tcaggtgtgc tgcattggtg
     1741 agttattttt tctgtctttg tggcaaataa tagacgtaaa aagtatttct aaaaaacaac
     1801 gaaaaacaaa attgttctgc ctatactttt tgaatttgaa ataaatattt ttcagctctt
     1861 cgggctcata ctttggtact cggtgccaaa aggtggttgg cacaaaccaa gcgacgccat
     1921 tcatccggta tcaaattacc cacccagaga cgaatttggg gaagaaaaat tattattttt
     1981 gacaggtgtg cctttatgga aggagctcgt caacatgctc gttcgccacc gccgttggct
     2041 tttttgccgt ctgtggagcc attgttctaa ttgttcttga cgccaagatg gaccaaatca
     2101 gttcggtgcc aacaagaagg agagctgtcc tagcagattt agttgtatcg ggtatttaat
     2161 ttgtgtgtgg aaaatttgaa agtaagagaa tttcagcaat tttcactgcc atatttttga
     2221 tcggtttttt cacattctgg tccaaattat ccgccttcga agtcgacgaa gacgacgaaa
     2281 atccaataaa gacaaacaac gcgaaattcg gaattctcag tgctcttctc tcctttttgg
     2341 cttgggtgcg tcatgaaaaa agtatagagg aatagggaaa acggagaggc atctgttatc
     2401 ttttgacaga atcttgagtt gaaaagcaac ctttttggcc actgttatcg agtgttacac
     2461 atcgtctttt aagtgtcaac gtgaaaggtt ggagagaatt tagtgaaata tgatataatt
     2521 tagtccatca gaaagagatt tgatactcaa gcggttttca gtagagcatc aaatagagaa
     2581 tacttgcaaa tcaattttgc acattttagg gaggtgccgc atttttcgca tggcgtcgtt
     2641 acgaagaagg aaaccaagca actcatgagc ccaattacga tgaacatttt ggacaagtgt
     2701 cgacagacgt gcaagatggt tatggatacg gaggagactc aaccggttag aataatttta
     2761 aacaaataga gaaaaaacta gttgaaaatt cttatgaaaa tcggatctca aattcagttt
     2821 gaaatttcaa aattctgtgc agaacacaac tactcaattt tttagttcac cactttttca
     2881 ttgtattctt atcacatttt gttcaacaaa tctttagaaa aacgttttac acgtattata
     2941 taatttttaa gtttatgact cttgagatat tacatgttaa aattgagcaa aatagcgtag
     3001 tactagtaga acaaaaaatt gatatatcaa atataccggt gaaaccacaa atacttcaaa
     3061 ctttgttaga agaatacaca aaaataatat ttacaataaa cttatctgaa acttcccagg
     3121 aagtttgttc actttttcaa aaaattatat ataagctagt tgaaacttgt tttacaattt
     3181 aaaaaaaagc aatttttttg agttttctct ggaagaaata tttttcataa gtgaatatat
     3241 gattttagtg gttttgagat tttctgcatt gaaaaaattc atttttctta acatttttta
     3301 gcaaaaactt ttaaaaaaat cacacgttgt ttttttcagg aatcggacat gttggcgcac
     3361 ctccacctca atcctcatac caatctggtg ctgcaccaca aaccatgcaa caaccaccat
     3421 caaacccata cactcagtcg gaaggatatg gttatggtgg cggagggtct ggtggcggag
     3481 ggtccggtgg cggagggtca ggggtaccga tgaagatgga caaaaagact atagtttggt
     3541 ttagaagaga cctaaggatt gaggataatc ctgcattagc agcagctgct cacgaaggat
     3601 ctgtttttcc tgtcttcatt tggtgtcctg aagaagaagg acagttttat cctggaagag
     3661 cttcaagatg gtggatgaaa caatcacttg ctcacttatc tcaatccttg aaggctcttg
     3721 gatctgacct cactttaatc aaaacccaca acacgatttc agcgatcttg gattgtatcc
     3781 gcgttaccgg tgctacaaaa gtcgtcttta accacctcta tgatcctgtt tcgttagttc
     3841 gggaccatac cgtaaaggag aagctggtgg aacgtgggat ctctgtgcaa agctacaatg
     3901 gagatctatt gtatgaaccg tgggagatat actgcgaaaa gggcaaacct tttacgagtt
     3961 tcaattctta ctggaagaaa tgcttagata tgtcgattga atccgttatg cttcctcctc
     4021 cttggcggtt gatgccaata actgcagcgg ctgaagcgat ttgggcgtgt tcgattgaag
     4081 aactagggct ggagaatgag gccgagaaac cgagcaatgc gttgttaact agagcttggt
     4141 ctccaggatg gagcaatgct gataagttac taaatgagtt catcgagaag cagttgatag
     4201 attatgcaaa gaacagcaag aaagttgttg ggaattctac ttcactactt tctccgtatc
     4261 tccatttcgg ggaaataagc gtcagacacg ttttccagtg tgcccggatg aaacaaatta
     4321 tatgggcaag agataagaac agtgaaggag aagaaagtgc agatcttttt cttaggggaa
     4381 tcggtttaag agagtattct cggtatatat gtttcaactt cccgtttact cacgagcaat
     4441 cgttgttgag tcatcttcgg tttttccctt gggatgctga tgttgataag ttcaaggcct
     4501 ggagacaagg caggaccggt tatccgttgg tggatgccgg aatgagagag ctttgggcta
     4561 ccggatggat gcataacaga ataagagtga ttgtttcaag ctttgctgtg aagtttcttc
     4621 tccttccatg gaaatgggga atgaagtatt tctgggatac acttttggct gctgatttgg
     4681 aatgtgacat ccttggctgg cagtatatct ctgggagtat ccccgatggc cacgagcttg
     4741 atcgcttgga caatcccgcg ttacaaggcg ccaaatatga cccagaaggt gagtacataa
     4801 ggcaatggct tcccgagctt gcgagattgc caactgaatg gatccatcat ccatgggacg
     4861 ctcctttaac cgtactcaaa gcttctggtg tggaactcgg aacaaactat gcgaaaccca
     4921 ttgtagacat cgacacagct cgtgagctac tagctaaagc tatttcaaga acccgtggag
     4981 cacagatcat gatcggagca gcacctgatg agattgtagc agatagcttc gaggccttag
     5041 gggctaatac cattaaagaa cctggtcttt gcccatctgt gtcttctaat gaccaacaag
     5101 taccttcggc tgtttaacct agggcctgca ggcccgctgt ctcatcctac tttcacctag
     5161 ttaactgctt gtcttaaaat ctatgcttct ctttagtatc taaaattttc ctagaagctt
     5221 acaagtatat aaatggtctc ttctcaataa aggttgtata tttattcatc ttattgaatc
     5281 tgccatttcc tcgtttttgc gagtttatat accttccaat tttctttcta ttgtattttc
     5341 aacttctaat tttaattcag ggaaactgct gtaccggtag aaaaaatgct cgagatggtg
     5401 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     5461 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     5521 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     5581 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     5641 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     5701 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     5761 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     5821 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     5881 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     5941 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     6001 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     6061 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagaatt ccaactgagc
     6121 gccggtcgct accattacca acttgtctgg tgtcaaaaat aataggggcc gctgtcatca
     6181 gagtaagttt aaactgagtt ctactaacta acgagtaata tttaaatttt cagcatctcg
     6241 cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc
     6301 cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt
     6361 tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa
     6421 ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc
     6481 caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt
     6541 tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta
     6601 tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat
     6661 cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta
     6721 attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc
     6781 gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg
     6841 tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg
     6901 tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt
     6961 tcctactagt cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7021 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7081 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     7141 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     7201 atgcgtaagg agaaaatacc gcatcaggcg gccttaatta agggcctcgt gatacgccta
     7261 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     7321 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7381 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     7441 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7501 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7561 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     7621 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     7681 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7741 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7801 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7861 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7921 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7981 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     8041 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     8101 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     8161 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     8221 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     8281 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     8341 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     8401 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     8461 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8521 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8581 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     8641 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8701 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8761 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8821 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8881 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8941 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     9001 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     9061 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     9121 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     9181 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     9241 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     9301 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     9361 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     9421 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctgtaagtt
     9481 taaacatgat cttactaact aactattctc atttaaattt tcagagctta aaaatggctg
     9541 aaatcactca caacgatgga tacgctaaca acttggaaat gaaataagct
//
