LOCUS       sequence-410735-        8464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SNG-1::CRY2olig(535) in GABAergic motor neurons of C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8464)
  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 8464)
  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: pDV18 [punc-47::SNG-1::CRY2olig(535)]
FEATURES             Location/Qualifiers
     source          1..8464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..25
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     22..38
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     22..38
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3517..5010
                     /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
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAA"
     primer_bind     complement(6071..6093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6193..6212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6290..6308)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6376..6480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6481..6549,6550..7341)
                     /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(6699..6718)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7512..8100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8001..8020
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8254..8271
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8388..8409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8424..8429,8430..8447,8448..8454)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(8462..8464,1..14)
                     /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)."
ORIGIN
        1 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctgtaag
       61 tttaaacatg atcttactaa ctaactattc tcatttaaat tttcagagct taaaaatggc
      121 tgaaatcact cacaacgatg gatacgctaa caacttggaa atgaaataag cttgcatgcc
      181 tgcagagctt gttgtcatca cttcaaactt ttcaatgcat aaatttcttg tcagatctga
      241 tcctctatac ttgaagttta taccgataaa ggttttcaac agattttgaa gcatttgaat
      301 caggggtgcc tgggtaaacc tatcatacga aactgccaat ttgtcctgtg aatcgtatct
      361 attaaaacaa gactgaaaat aactaaactt ctacgtcaaa aagttgacaa aacaactttc
      421 ttggaaaaaa agctgttggg atcccggaac agtcgaaagt cggtggcaag cgccgaactg
      481 ctgacggtct aaccggggca caaatcaggg gtgagcggca aacgattttt ccggcaaatc
      541 ggcaaatcgg caaattgcca atattgaaat acccggcaaa tcggtaaata gccggaattg
      601 aaaatttccg gcaaactggt aaaccgcaaa ttgctgattt gccgaatttg ccgggaagac
      661 ggcaattgcc aaacatattc ggcaaattgt ggttttgcac tttttggaaa tttcagaatt
      721 tcaatcggca aattgtgcac atcctatgaa ttttcctacg tctattttga aaagtaagca
      781 aattctatga aaatatctaa agaaaaatgg aaaaaatttt caaaaaggca cagttttaag
      841 tgtttccgtc taataaaaaa atccccctaa acacttccgg caaattgatg ttcggcaaat
      901 ggcaaatcgg aaacttgccg aaaattacag tttccggtaa atcggcaaac cggcaaactg
      961 cctgaattga aaagttccgt caaatcggca aaccgacaac acccctggca caaatgatgg
     1021 acatactgag gcaatttgcc ggttttccaa ttgcaggaaa ttttcaattc cggcagtgtg
     1081 ccgatttgcc ggaaatttta attcaggcaa attgccgatt tcccgatttc ccgatttgcc
     1141 ggaaaaaatc gtttgccgcc cacccctggg tctgaacctt gattgttaca aaacattttt
     1201 agctctttgg agaaataaaa tgaatctcgt aaaatttaat tgacgaggac gatattagct
     1261 gtctctttag accaaattca gaaaaaaaga aagaatactt cccaaatttc cggtccctct
     1321 ctcgtttttt ttgccaataa actcactata gtcgctggtt cccccctatt cacatttatt
     1381 ctaccaatcc atcagtggaa ccagaaaaaa gaagagcctt tcggtttgga gagtagggtc
     1441 taataatccc ccgtgctctt caaatcattg tgccaacaca cagacacact ttatgtgtgc
     1501 tcacacacac acgctatttg aagagcgaag acgacgacga cgcattcaga gctcttttcc
     1561 acgaaatttg ctccatcttt ccacaatctg tctttcctgt gagacgacag cgtcacattt
     1621 atttcattac agcaccggtg aaatggagaa cgtgcgtgct tatggtgccg gactagctgg
     1681 cgccaatttc gacaagaaca catttttcaa aaaaccaacg gttttgttca ggtgtgctgc
     1741 attggtgagt tattttttct gtctttgtgg caaataatag acgtaaaaag tatttctaaa
     1801 aaacaacgaa aaacaaaatt gttctgccta tactttttga atttgaaata aatatttttc
     1861 agctcttcgg gctcatactt tggtactcgg tgccaaaagg tggttggcac aaaccaagcg
     1921 acgccattca tccggtatca aattacccac ccagagacga atttggggaa gaaaaattat
     1981 tatttttgac aggtgtgcct ttatggaagg agctcgtcaa catgctcgtt cgccaccgcc
     2041 gttggctttt ttgccgtctg tggagccatt gttctaattg ttcttgacgc caagatggac
     2101 caaatcagtt cggtgccaac aagaaggaga gctgtcctag cagatttagt tgtatcgggt
     2161 atttaatttg tgtgtggaaa atttgaaagt aagagaattt cagcaatttt cactgccata
     2221 tttttgatcg gttttttcac attctggtcc aaattatccg ccttcgaagt cgacgaagac
     2281 gacgaaaatc caataaagac aaacaacgcg aaattcggaa ttctcagtgc tcttctctcc
     2341 tttttggctt gggtgcgtca tgaaaaaagt atagaggaat agggaaaacg gagaggcatc
     2401 tgttatcttt tgacagaatc ttgagttgaa aagcaacctt tttggccact gttatcgagt
     2461 gttacacatc gtcttttaag tgtcaacgtg aaaggttgga gagaatttag tgaaatatga
     2521 tataatttag tccatcagaa agagatttga tactcaagcg gttttcagta gagcatcaaa
     2581 tagagaatac ttgcaaatca attttgcaca ttttagggag gtgccgcatt tttcgcatgg
     2641 cgtcgttacg aagaaggaaa ccaagcaact catgagccca attacgatga acattttgga
     2701 caagtgtcga cagacgtgca agatggttat ggatacggag gagactcaac cggttagaat
     2761 aattttaaac aaatagagaa aaaactagtt gaaaattctt atgaaaatcg gatctcaaat
     2821 tcagtttgaa atttcaaaat tctgtgcaga acacaactac tcaatttttt agttcaccac
     2881 tttttcattg tattcttatc acattttgtt caacaaatct ttagaaaaac gttttacacg
     2941 tattatataa tttttaagtt tatgactctt gagatattac atgttaaaat tgagcaaaat
     3001 agcgtagtac tagtagaaca aaaaattgat atatcaaata taccggtgaa accacaaata
     3061 cttcaaactt tgttagaaga atacacaaaa ataatattta caataaactt atctgaaact
     3121 tcccaggaag tttgttcact ttttcaaaaa attatatata agctagttga aacttgtttt
     3181 acaatttaaa aaaaagcaat ttttttgagt tttctctgga agaaatattt ttcataagtg
     3241 aatatatgat tttagtggtt ttgagatttt ctgcattgaa aaaattcatt tttcttaaca
     3301 ttttttagca aaaactttta aaaaaatcac acgttgtttt tttcaggaat cggacatgtt
     3361 ggcgcacctc cacctcaatc ctcataccaa tctggtgctg caccacaaac catgcaacaa
     3421 ccaccatcaa acccatacac tcagtcggaa ggatatggtt atggtggcgg agggtctggt
     3481 ggcggagggt ccggtggcgg agggtcaggg gtaccgatga agatggacaa aaagactata
     3541 gtttggttta gaagagacct aaggattgag gataatcctg cattagcagc agctgctcac
     3601 gaaggatctg tttttcctgt cttcatttgg tgtcctgaag aagaaggaca gttttatcct
     3661 ggaagagctt caagatggtg gatgaaacaa tcacttgctc acttatctca atccttgaag
     3721 gctcttggat ctgacctcac tttaatcaaa acccacaaca cgatttcagc gatcttggat
     3781 tgtatccgcg ttaccggtgc tacaaaagtc gtctttaacc acctctatga tcctgtttcg
     3841 ttagttcggg accataccgt aaaggagaag ctggtggaac gtgggatctc tgtgcaaagc
     3901 tacaatggag atctattgta tgaaccgtgg gagatatact gcgaaaaggg caaacctttt
     3961 acgagtttca attcttactg gaagaaatgc ttagatatgt cgattgaatc cgttatgctt
     4021 cctcctcctt ggcggttgat gccaataact gcagcggctg aagcgatttg ggcgtgttcg
     4081 attgaagaac tagggctgga gaatgaggcc gagaaaccga gcaatgcgtt gttaactaga
     4141 gcttggtctc caggatggag caatgctgat aagttactaa atgagttcat cgagaagcag
     4201 ttgatagatt atgcaaagaa cagcaagaaa gttgttggga attctacttc actactttct
     4261 ccgtatctcc atttcgggga aataagcgtc agacacgttt tccagtgtgc ccggatgaaa
     4321 caaattatat gggcaagaga taagaacagt gaaggagaag aaagtgcaga tctttttctt
     4381 aggggaatcg gtttaagaga gtattctcgg tatatatgtt tcaacttccc gtttactcac
     4441 gagcaatcgt tgttgagtca tcttcggttt ttcccttggg atgctgatgt tgataagttc
     4501 aaggcctgga gacaaggcag gaccggttat ccgttggtgg atgccggaat gagagagctt
     4561 tgggctaccg gatggatgca taacagaata agagtgattg tttcaagctt tgctgtgaag
     4621 tttcttctcc ttccatggaa atggggaatg aagtatttct gggatacact tttggatgct
     4681 gatttggaat gtgacatcct tggctggcag tatatctctg ggagtatccc cgatggccac
     4741 gagcttgatc gcttggacaa tcccgcgtta caaggcgcca aatatgaccc agaaggtgag
     4801 tacataaggc aatggcttcc cgagcttgcg agattgccaa ctgaatggat ccatcatcca
     4861 tgggacgctc ctttaaccgt actcaaagct tctggtgtgg aactcggaac aaactatgcg
     4921 aaacccattg tagacatcga cacagctcgt gagctactag ctaaagctat ttcaagaacc
     4981 cgtggagcac agatcatgat cggagcagca cctgatgaga ttgtagcaga tagcttcgag
     5041 gccttagggg ctaataccat taaagaacct ggtctttgcc catctgtgtc ttctaatgac
     5101 caacaagtac cttcggctgt ttaacctagg gcctgcaggc ccgggagcat tcgtagaatt
     5161 ccaactgagc gccggtcgct accattacca acttgtctgg tgtcaaaaat aataggggcc
     5221 gctgtcatca gagtaagttt aaactgagtt ctactaacta acgagtaata tttaaatttt
     5281 cagcatctcg cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat
     5341 gctctttctc cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat
     5401 ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa
     5461 atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat
     5521 aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga
     5581 taaatttttt tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg
     5641 tttcagttta tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa
     5701 tcatgctcat cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt
     5761 tatgaaatta attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt
     5821 cgaggacggc gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat
     5881 cggaagaagg tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag
     5941 tggagtagtg tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa
     6001 cataactgtt tcctactagt cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg
     6061 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     6121 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     6181 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     6241 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg cctcgtgata
     6301 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     6361 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6421 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6481 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6541 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6601 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6661 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6721 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6781 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6841 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6901 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6961 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7021 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7081 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7141 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7201 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7261 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7321 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7381 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7441 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7501 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7561 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7621 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7681 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7741 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7801 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7861 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7921 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7981 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8041 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8101 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8161 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8221 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8281 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     8341 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     8401 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     8461 attg
//
