LOCUS       sequence-360435-        7628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for CK(0.4)::Opto-vTrap.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7628)
  AUTHORS   Won J, Pankratov Y, Jang MW, Kim S, Ju YH, Lee S, Lee SE, Kim A, 
            Park S, Lee CJ, Heo WD
  TITLE     Opto-vTrap, an optogenetic trap for reversible inhibition of 
            vesicular release, synaptic transmission, and behavior.
  JOURNAL   Neuron. 2022 Feb 2;110(3):423-435.e4. doi: 
            10.1016/j.neuron.2021.11.003. Epub 2021 Nov 30.
  PUBMED    34852235
REFERENCE   2  (bases 1 to 7628)
  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"; doi: 
            "10.1016/j.neuron.2021.11.003"; journalName: "Neuron"; date: 
            "2022-02-2- 2"; volume: "110"; issue: "3"; pages: "423-435.e4"
COMMENT     Sequence Label: pAAV-CK(0.4)-CRY2PHR-P2A-CIBN-mCherry-VAMP2
FEATURES             Location/Qualifiers
     source          1..7628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             2041..2097
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2098..2607
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     regulatory      2620..2629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2626..3333
                     /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(2771..2789)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3039..3058
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3755..4343
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3808..3828)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4226..4237)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4375..4851
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4574..4593)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4891..5031
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4891..5020
                     /label=AAV2 ITR
     rep_origin      5106..5561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5193..5212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5403..5424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5578..5597)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5697..5719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5757..5775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5843..5947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5948..6016,6017..6808)
                     /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(6166..6185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6979..7567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7468..7487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtacttgt ggactaagtt tgttcacatc cccttctcca
      181 accccctcag tacatcaccc tgggggaaca gggtccactt gctcctgggc ccacacagtc
      241 ctgcagtatt gtgtatataa ggccagggca aagaggagca ggttttaaag tgaaaggcag
      301 gcaggtgttg gggaggcagt taccggggca acgggaacag ggcgtttcgg aggtggttgc
      361 catggggacc tggatgctga cgaaggctcg cgaggctgtg agcagccaca gtgccctgct
      421 cagaagcccc aagctcgtca gtcaagccgg ttctccgttt gcactcagga gcacgggcag
      481 gcgagtggcc cctagttctg ggggcagcgg atccttcgaa accggtgcta gcccaccatg
      541 aagatggaca aaaagaccat cgtctggttt cggagagatt tgagaataga agataatccc
      601 gcgctcgccg ccgcggccca cgagggttcc gtcttccccg ttttcatttg gtgtcctgaa
      661 gaagaaggcc agttttatcc cggaagggcc tctaggtggt ggatgaagca aagtctggcc
      721 catcttagcc agtcactgaa agcactgggc agtgatctta ccctgatcaa gacacacaat
      781 accatctctg ccatcctcga ctgcatcagg gtgaccggcg caacgaaagt cgtgtttaac
      841 cacctgtacg atccagttag tctggtgcgc gaccacactg tgaaggagaa gctggtggaa
      901 cgggggatca gtgtgcagag ctacaacggg gaccttctgt acgagccatg ggagatctat
      961 tgcgagaaag ggaaaccgtt cacctccttc aacagttact ggaagaaatg tttggatatg
     1021 tcaatagagt ccgttatgtt gccccctccc tggagactga tgccgattac tgctgctgca
     1081 gaggccatct gggcctgctc catcgaggaa ctcggtctgg aaaatgaagc agaaaagcca
     1141 agcaatgcac ttctcactag agcctggagc cccggctggt ctaatgccga caagctgctt
     1201 aacgagttca tcgaaaaaca actgattgac tacgcgaaga actccaagaa agtggtaggt
     1261 aactcaacta gcttgctctc tccatatctc cattttggcg agatttctgt ccgccatgta
     1321 tttcagtgcg ctcggatgaa acagattatc tgggctcgcg ataaaaacag cgaaggcgaa
     1381 gaaagcgccg atctgttcct gcgagggatc ggacttcggg aatactcccg gtatatatgt
     1441 ttcaactttc cattcacaca cgagcagagt ctgttgtccc acctcaggtt cttcccctgg
     1501 gacgccgatg tcgacaaatt caaggcatgg agacagggaa ggacagggta cccactcgtg
     1561 gatgctggca tgagagagct ctgggctaca ggctggatgc acaaccgcat ccgggtaatc
     1621 gtgtcctcat ttgctgtcaa gtttctgctc ctgccttgga aatggggaat gaagtacttt
     1681 tgggataccc ttctcgacgc cgacttggag tgtgacattc tgggatggca atatattagc
     1741 gggtcaattc ctgacggcca tgagttggac aggttggaca atccggcctt gcagggagct
     1801 aagtatgatc ccgaaggaga gtatattcga cagtggctcc ccgagctggc ccgacttcct
     1861 acggagtgga ttcaccatcc ttgggacgca ccactgacag tgctcaaggc aagcggggtg
     1921 gagctgggca ccaattacgc taagcctata gttgatatag atacagcacg cgagctgctg
     1981 gctaaagcga tctctcgcac tcgggaggcg cagattatga tcggtgctgc cggcagcggc
     2041 gccacgaact tctctctgtt aaagcaagca ggagacgtgg aagaaaaccc cggtcctatg
     2101 aatggagcta taggaggtga ccttttgctc aattttcctg acatgtcggt cctagagcgc
     2161 caaagggctc acctcaagta cctcaatccc acctttgatt ctcctctcgc cggcttcttt
     2221 gccgattctt caatgattac cggcggcgag atggacagct atctttcgac tgccggtttg
     2281 aatcttccga tgatgtacgg tgagacgacg gtggaaggtg attcaagact ctcaatttcg
     2341 ccggaaacga cgcttgggac tggaaatttc aaggcagcga agtttgatac agagactaag
     2401 gattgtaatg aggcggcgaa gaagatgacg atgaacagag atgacctagt agaagaagga
     2461 gaagaagaga agtcgaaaat aacagagcaa aacaatggga gcacaaaaag catcaagaag
     2521 atgaaacaca aagccaagaa agaagagaac aatttctcta atgattcatc taaagtgacg
     2581 aaggaattgg agaaaacgga ttatattcat gtaccggtcg ccaccatggt gagcaagggc
     2641 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2701 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2761 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2821 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2881 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2941 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     3001 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     3061 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3121 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     3181 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     3241 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3301 cactccaccg gcggcatgga cgagctgtac aagggcagtg gttccggact cagatctcga
     3361 gctatgtcgg ctaccgctgc caccgtcccg cctgccgccc cggccggcga gggtggcccc
     3421 cctgcacctc ctccaaacct tactagtaac aggagactgc agcagaccca ggcccaggtg
     3481 gatgaggtgg tggacatcat gagggtgaat gtggacaagg tcctggagcg ggaccagaag
     3541 ttgtcggagc tggatgaccg tgcagatgcc ctccaggcag gggcctccca gtttgaaaca
     3601 agtgcagcca agctcaagcg caaatactgg tggaaaaacc tcaagatgat gatcatcttg
     3661 ggagtgatct gcgccatcat cctcatcatc atcatagttt acttcagctc ttaaggcgcg
     3721 cccctgcagg gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     3781 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3841 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3901 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3961 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4021 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4081 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4141 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     4201 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4261 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4321 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     4381 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4441 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4501 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4561 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     4621 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     4681 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     4741 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     4801 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     4861 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     4921 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4981 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     5041 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5101 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5161 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     5221 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5281 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5341 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5401 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     5461 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     5521 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5581 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5641 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5701 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5761 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5821 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5881 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5941 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6001 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6061 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6121 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6181 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6241 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6301 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6361 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6421 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6481 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6541 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6601 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6661 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6721 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6781 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6841 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6901 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6961 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7021 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7081 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7141 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7201 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7261 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7321 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7381 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7441 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7501 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7561 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7621 tcacatgt
//
