LOCUS       sequence-379301-        5494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for optogenetic long-term silencing of excitatory 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5494)
  AUTHORS   Wiegert Lab
  TITLE     Wiegert Lab Plasmids 2022
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5494)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CaMKII-Aion-ts-citrine-ER
FEATURES             Location/Qualifiers
     source          1..5494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      510..519
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1518..1520,1521..1523,1524..2234)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1563..1584)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1824..1843)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2171..2192
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2265..2668
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    2675..2723
                     /note="synthetic polyadenylation signal"
     repeat_region   2753..2893
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2753..2882
                     /label=AAV2 ITR
     rep_origin      2968..3423
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3055..3074)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3265..3286
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3440..3459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3559..3581
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3619..3637)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3705..3809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3810..3878,3879..4670)
                     /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(4028..4047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4841..5429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5330..5349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5491..5494,1..126)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc
       61 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt cttgtggact aagtttgttc gcatcccctt ctccaacccc
      181 ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca cagtcctgca
      241 gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa ggcaggcagg
      301 tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg gttgccatgg
      361 ggacctggat gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc ctgctcagaa
      421 gccccaagct cgtcagtcaa gccggttctc cgtttgcact caggagcacg ggcaggcgag
      481 tggcccctag ttctgggggc agctctagcg ccaccatgga ctatggcggc gctttgtctg
      541 ccgtcggact cttccaaacc tcatacactc ttgagaacaa tggttctgtt atttgcatcc
      601 cgaacaacgg ccagtgcttc tgcttggctt ggcttaaatc caacggaaca aatgccgaga
      661 agttggctgc caacattctg cagtggattt cttttgcgct ttcagcgctc tgcctgatgt
      721 tctacggcta ccagacctgg aagtctactt gcggctggga gaacatttac gtggccacga
      781 tccagatgat caagttcatc atcgagtatt tccatagctt tgacgaacct gcggtgatct
      841 actcatccaa cggcaacaag acccggtggc ttcgttacgc gagctggctg ctgaccgccc
      901 ctgtcattct tatccacctg agcaacctta cgggtctggc gaacgactat aacaagcgta
      961 ccatgggtct gctggtgtca tgcatcggcg gcatcgtgtg ggccaccacg gccgcgctgt
     1021 ccaagggata cgtccgtgtc attttcttcc tgatgggcct gtgctacggc atctacacat
     1081 tcttcaacgc agccaaggtc tacattgagg cgtaccacac cgtgccgaag ggccggtgtc
     1141 gccaggtggt gactggcatg gcttggctct tcttcgtatc atggggtatg ttccccatcc
     1201 tgttcatcct cggccccgag ggcttcggcg tcctgagccg gtacggctcc aacgtcggcc
     1261 acaccatcat tgacctgatg tcgaagcagt gctggggtct gctcggccac tacctgcgcg
     1321 tgctgatcca cagccatatc ctcatccacg gcgacattcg caagaccacc aaattgaaca
     1381 ttggtggcac tgagattgag gtcgagacgc tggtggagga cgaggccgag gctggcgcgg
     1441 tcgcggcact cgagaagagc aggatcacca gcgagggcga gtacatcccc ctggaccaga
     1501 tcgacatcaa cgtggtgatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1561 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1621 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1681 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct
     1741 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1801 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1861 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1921 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1981 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2041 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2101 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     2161 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2221 acgagctgta caagttctgc tacgagaacg aggtgtaaaa gcttaatcaa cctctggatt
     2281 acaaaatttg tgaaagattg actgatattc ttaactatgt tgctcctttt acgctgtgtg
     2341 gatatgctgc tttaatgcct ctgtatcatg ctattgcttc ccgtatggct ttcgttttct
     2401 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtccgtc
     2461 aacgtggcgt ggtgtgctct gtgtttgctg acgcaacccc cactggctgg ggcattgcca
     2521 ccacctgtca actcctttct gggactttcg ctttccccct cccgatcgcc acggcagaac
     2581 tcatcgccgc ctgccttgcc cgctgctgga caggggctag gttgctgggc actgataatt
     2641 ccgtggtgtt gtcggggaag ctgacgtcag cgctaataaa atcgctatcc atcgaagatg
     2701 gatgtgtgtt ggttttttgt gtgggtaacc cacgtgcgga ccgagcggcc gcaggaaccc
     2761 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2821 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2881 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2941 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3001 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     3061 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3121 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3181 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3241 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     3301 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     3361 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3421 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     3481 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     3541 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     3601 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3661 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3721 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3781 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3841 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3901 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3961 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4021 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4081 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4141 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4201 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4261 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4321 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4381 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4441 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4501 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4561 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4621 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4681 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5341 ggtatcttta tagtcctgtc gggttttgcc acctctgact tgagcgtcga tttttgtgat
     5401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5461 tggccttttg ctggcctttt gctcacatgt cctg
//
