LOCUS       sequence-379280-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for bidirectional optogenetic manipulation of excitatory 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  AUTHORS   Wiegert Lab
  TITLE     Wiegert Lab Plasmids 2022
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7044)
  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            "Unpublished"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-somBiPOLES-eGFP
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(1490..1492,1493..2203)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1490..1492,1493..2203)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1532..1553)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1793..1812)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2140..2161
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2564..3607
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=Chrimson
                     /translation="ELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTEWW
                     CDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSIAI
                     ALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRYFE
                     WLLSCPVILIKLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIVSC
                     IYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLKLS
                     PYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEEFV
                     EEEDEDTV"
     misc_feature    3824..4227
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    4234..4282
                     /note="synthetic polyadenylation signal"
     repeat_region   4312..4452
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4312..4441
                     /label=AAV2 ITR
     rep_origin      4527..4982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4614..4633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4824..4845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4999..5018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5118..5140
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5178..5196)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5264..5368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5369..5437,5438..6229)
                     /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(5587..5606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6400..6988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6889..6908
                     /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 cgcacgcgtc ttgtggacta agtttgttcg catccccttc
      181 tccaaccccc tcagtacatc accctggggg aacagggtcc acttgctcct gggcccacac
      241 agtcctgcag tattgtgtat ataaggccag ggcaaagagg agcaggtttt aaagtgaaag
      301 gcaggcaggt gttggggagg cagttaccgg ggcaacggga acagggcgtt tcggaggtgg
      361 ttgccatggg gacctggatg ctgacgaagg ctcgcgaggc tgtgagcagc cacagtgccc
      421 tgctcagaag ccccaagctc gtcagtcaag ccggttctcc gtttgcactc aggagcacgg
      481 gcaggcgagt ggcccctagt tctgggggca gctctagagc ggtaccggat ccatggcatc
      541 acaggtcgtc tatggggaat gggcatcaac tcacaccgag tgctacaata tgtccagaat
      601 cgactccacc ttcgtctcac tgctgcagct ggtgtgggcc gtggtctcag gctgccagac
      661 aatcttcatg attagcaggg ctcccaaggt cccttgggag tccgtgtacc tgccctttgt
      721 cgaatctatc acttatgcac tggccagtac tggcaacgga accctgcaga tgagggacgg
      781 cagattcttt ccttggagcc gaatggcctc ctggctgtgc acctgtccaa tcatgctggg
      841 acagatttct aacatggctc tggtgaagta caaaagtatc ccactgaatc ccattgcaca
      901 ggccgcttcc atcattagag tggtcatggg aatcaccgct acaatttctc ccgcagagta
      961 tatgaagtgg ctgttctttt tctttggggc cacctgcctg gtgttcgagt acagtgtggt
     1021 cttcacaatc tttcaagtgg ggctgtatgg ctttgaatct gtcgggacac ctctggcaca
     1081 gaaggtggtg gtgaggatca agatgctgcg cctgatcttc tttattgctt ggactatgtt
     1141 cccaatcgtg tggctgattt cacccaccgg agtctgcgtg atccacgaga acgtgagcgc
     1201 cattctgtac ctgctggctg acggcctgtg caaaaataca tatggagtga tcctgtggag
     1261 cactgcctgg ggagtgctgg aagggaagtg ggaccccgca tgtctgccag ggcaggagaa
     1321 acccgaagcc gacgatcctt ttggcctgaa tcacgagaag aatgcccctc ctaacgacga
     1381 agtcaatatc agaatgtttg gaaggggacg tacggcggcc aagagcagga tcaccagcga
     1441 gggcgagtac atccccctgg accagatcga catcaacgtg gcggtacccg tgagcaaggg
     1501 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1561 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1621 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1681 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1741 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1801 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1861 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1921 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1981 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2041 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2101 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2161 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagaccggtg cagccctgca
     2221 ggagaagaag tcatgcagcc agcgcatggc cgaattccgg caatactgtt ggaacccgga
     2281 cactgggcag atgctgggcc gcaccccagc ccggtgggtg tggatcagcc tgtactatgc
     2341 agctttctac gtggtcatga ctgggctctt tgccttgtgc atctatgtgc tgatgcagac
     2401 cattgatccc tacacccccg actaccagga ccagttaaag tcaccggggg taaccttgag
     2461 accggatgtg tatggggaaa gagggctgca gatttcctac aacatctctg aaaacagctc
     2521 tagacaggcc cagatcaccg gacgtccgga gggagctgta caagagctga tcagcagcgc
     2581 caccagatct ctgtttgccg ccggaggcat caacccttgg cctaacccct accaccacga
     2641 ggacatgggc tgtggaggaa tgacacctac aggcgagtgc ttcagcaccg agtggtggtg
     2701 tgacccttct tacggactga gcgacgccgg atacggatat tgcttcgtgg aggccacagg
     2761 cggctacctg gtcgtgggag tggagaagaa gcaggcttgg ctgcacagca gaggcacacc
     2821 aggagaaaag atcggcgccc aggtctgcca gtggattgct ttcagcatcg ccatcgccct
     2881 gctgacattc tacggcttca gcgcttggaa ggccacttgc ggttgggagg aggtctacgt
     2941 ctgttgcgtc gaggtgctgt tcgtgaccct ggagatcttc aaggagttca gcagccccgc
     3001 cacagtgtac ctgtctaccg gcaaccacgc ctattgcctg cgctacttcg agtggctgct
     3061 gtcttgcccc gtgatcctga tcaagctgag caacctgagc ggcctgaaga acgactacag
     3121 caagcggacc atgggcctga tcgtgtcttg cgtgggaatg atcgtgttcg gcatggccgc
     3181 aggactggct accgattggc tcaagtggct gctgtatatc gtgtcttgca tctacggcgg
     3241 ctacatgtac ttccaggccg ccaagtgcta cgtggaagcc aaccacagcg tgcctaaagg
     3301 ccattgccgc atggtcgtga agctgatggc ctacgcttac ttcgcctctt ggggcagcta
     3361 cccaatcctc tgggcagtgg gaccagaagg actgctgaag ctgagccctt acgccaacag
     3421 catcggccac agcatctgcg acatcatcgc caaggagttt tggaccttcc tggcccacca
     3481 cctgaggatc aagatccacg agcacatcct gatccacggc gacatccgga agaccaccaa
     3541 gatggagatc ggaggcgagg aggtggaagt ggaagagttc gtggaggagg aggacgagga
     3601 cacagtgctc gagcagagcc agcccatcct gaacaccaag gagatggccc cccagagcaa
     3661 gccccccgag gagctggaga tgagcagcat gcccagcccc gtggcccccc tgcccgcccg
     3721 caccgagggc gtgatcgaca tgcgcagcat gagcagcatc gacagcttca tcagctgcgc
     3781 caccgacttc cccgaggcca cccgcttctg atctagaaag cttaatcaac ctctggatta
     3841 caaaatttgt gaaagattga ctgatattct taactatgtt gctcctttta cgctgtgtgg
     3901 atatgctgct ttaatgcctc tgtatcatgc tattgcttcc cgtatggctt tcgttttctc
     3961 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtccgtca
     4021 acgtggcgtg gtgtgctctg tgtttgctga cgcaaccccc actggctggg gcattgccac
     4081 cacctgtcaa ctcctttctg ggactttcgc tttccccctc ccgatcgcca cggcagaact
     4141 catcgccgcc tgccttgccc gctgctggac aggggctagg ttgctgggca ctgataattc
     4201 cgtggtgttg tcggggaagc tgacgtcagc gctaataaaa tcgctatcca tcgaagatgg
     4261 atgtgtgttg gttttttgtg tgggtaaccc acgtgcggac cgagcggccg caggaacccc
     4321 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4381 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     4441 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4501 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     4561 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     4621 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4681 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4741 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4801 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     4861 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     4921 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4981 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     5041 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5101 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5161 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5221 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5281 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5341 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5401 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5461 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5521 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5581 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5641 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5701 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5761 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5821 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5881 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5941 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     6001 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6061 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6121 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6181 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6241 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6301 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6541 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6601 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6661 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6721 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6781 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6841 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6901 gtatctttat agtcctgtcg ggttttgcca cctctgactt gagcgtcgat ttttgtgatg
     6961 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7021 ggccttttgc tggccttttg ctca
//
