LOCUS       sequence-256083-        7110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of ChRmine-eYFP under the control
            of CamKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7110)
  AUTHORS   Marshel JH, Kim YS, Machado TA, Quirin S, Benson B, Kadmon J, Raja 
            C, Chibukhchyan A, Ramakrishnan C, Inoue M, Shane JC, McKnight DJ, 
            Yoshizawa S, Kato HE, Ganguli S, Deisseroth K
  TITLE     Cortical layer-specific critical dynamics triggering perception.
  JOURNAL   Science. 2019 Aug 9;365(6453). pii: science.aaw5202. doi: 
            10.1126/science.aaw5202. Epub 2019 Jul 18.
  PUBMED    31320556
REFERENCE   2  (bases 1 to 7110)
  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: "Science."; 
            date: "2019-08-9"; volume: "365(6453). pii: science.aaw5202. doi"; 
            pages: " 10.1126/science.aaw5202. Epub 2019 Jul 18"
COMMENT     Sequence Label: pAAV-CaMKIIa-ChRmine-eYFP-WPRE
FEATURES             Location/Qualifiers
     source          1..7110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        154..1442
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1475..1484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2477..2479,2480..3190)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2519..2540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2780..2799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3127..3148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3242..3830
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3295..3315)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3713..3724)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3862..4338
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4061..4080)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4378..4518
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4378..4507
                     /label=AAV2 ITR
     rep_origin      4593..5048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4680..4699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4890..4911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5065..5084)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5184..5206
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5244..5262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5330..5434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5435..5503,5504..6295)
                     /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(5653..5672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6466..7054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6955..6974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtt taacattatg gccttaggtc acttcatctc
      181 catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga gcttcctcag
      241 tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca gattactaat
      301 cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg aactcattag
      361 ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc tacaccaccc
      421 gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct ctccagtcct
      481 accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc aaatcttcat
      541 ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag gggccctgct
      601 gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca gcattctggg
      661 tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat ggctaaaaag
      721 cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca gaccaattcc
      781 cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga gagatgaatt
      841 agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca atacctgtct
      901 cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag gaccatccca
      961 gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac ttaatcccat
     1021 cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag cgatgatcac
     1081 ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc accctggggg
     1141 aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat ataaggccag
     1201 ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg
     1261 ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg
     1321 ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag
     1381 ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca
     1441 gctctagagc ggtaccggat ccccgggtac cggtgccacc atggcacacg caccaggcac
     1501 cgaccagatg ttctacgtgg gcacaatgga cggctggtat ctggatacca agctgaactc
     1561 cgtggccatc ggcgcccact ggtcttgctt tatcgtgctg acaatcacca cattctacct
     1621 gggctatgag tcctggacat ctcggggccc tagcaagaga acctcctttt acgccggcta
     1681 tcaggaggag cagaacctgg ccctgtttgt gaatttcttt gccatgctga gctacttcgg
     1741 caagatcgtg gccgacacac tgggccacaa cttcggcgat gtgggcccct tcatcatcgg
     1801 cttcggcaat tacaggtatg ccgactacat gctgacctgc ccaatgctgg tgtacgatct
     1861 gctgtatcag ctgagggccc cctatcgcgt gtcttgtagc gccatcatct ttgccatcct
     1921 gatgtctggc gtgctggccg agttctacgc agagggcgac cctaggctga ggaatggcgc
     1981 ctacgcctgg tatggctttg gctgtttctg gtttatcttc gcctactcta tcgtgatgag
     2041 catcgtggcc aagcagtata gccggctggc ccagctggcc caggatacag gcgccgagca
     2101 cagcctgcac gtgctgaagt tcgccgtgtt taccttctcc atgctgtgga ttctgtttcc
     2161 cctggtgtgg gccatctgcc ctagaggctt cggctggatc gacgataact ggacagaggt
     2221 ggcccactgc gtgtgcgaca tcgtggccaa gtcctgttac ggctttgccc tggcccggtt
     2281 cagaaagacc tatgatgagg agctgtttcg gctgctggag cagctgggac acgacgagga
     2341 tgagttccag aagctggagc tggatatgag gctgagcagc aatggcgagc gcctgcggag
     2401 actgtctgcg gccgccaaga gcaggatcac cagcgagggc gagtacatcc ccctggacca
     2461 gatcgacatc aacgtggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2521 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2581 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2641 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
     2701 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2761 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2821 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2881 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2941 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3001 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3061 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     3121 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3181 gctgtacaag ttctgctacg agaacgaggt gtaatgagaa ttcgatatca agcttatcga
     3241 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3301 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3361 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3421 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3481 tggttgggga attgccacca cctgtcagct cctttccgga actttcgctt tccccctccc
     3541 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3601 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3661 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3721 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3781 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3841 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     3901 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     3961 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     4021 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     4081 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     4141 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     4201 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     4261 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     4321 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     4381 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4441 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4501 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4561 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     4621 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     4681 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4741 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4801 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4861 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4921 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     4981 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     5041 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     5101 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5161 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5221 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5281 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5341 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5401 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5461 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5521 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5581 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5641 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5701 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5761 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5821 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5881 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5941 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6001 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6061 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6121 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6181 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6241 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6301 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6361 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6421 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6481 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6541 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6601 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6661 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6721 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6781 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6841 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6901 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6961 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7021 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7081 gttcctggcc ttttgctggc cttttgctca
//
