LOCUS       sequence-385172-        7875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV2 transfer plasmid for iLMO2, an opto-chemogenetic probe for 
            neuronal inhibition by light or chemical, under control of the 
            glutamatergic-selective CaMKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7875)
  AUTHORS   Tung JK, Gutekunst CA, Gross RE
  TITLE     Inhibitory luminopsins: genetically-encoded bioluminescent opsins 
            for versatile, scalable, and hardware-independent optogenetic 
            inhibition.
  JOURNAL   Sci Rep. 2015 Sep 24;5:14366. doi: 10.1038/srep14366.
  PUBMED    26399324
REFERENCE   2  (bases 1 to 7875)
  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: "Sci Rep."; 
            date: "2015-09-24"; volume: "5:14366. doi"; pages: " 
            10.1038/srep14366"
COMMENT     Sequence Label: pAAV/CaMKIIa::iLMO2
FEATURES             Location/Qualifiers
     source          1..7875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             join(2340..2342,2343..2855)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(2382..2403)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2643..2662)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2990..3011
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4002..4590
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4055..4075)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4473..4484)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4622..5098
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4821..4840)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5138..5278
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5138..5267
                     /label=AAV2 ITR
     rep_origin      5353..5808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5440..5459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5650..5671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5825..5844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5944..5966
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6004..6022)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6090..6194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6195..6263,6264..7055)
                     /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(6413..6432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7226..7814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7715..7734
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccatg acagagaccc tgcctcccgt gaccgagagt gccgtggccc
     1501 ttcaagccga ggttacccaa agggagttgt tcgagttcgt gctgaacgac cctttgcttg
     1561 caagcagtct ctatatcaac atcgcacttg caggactgag tatactgctg ttcgttttta
     1621 tgacccgagg actcgatgat ccacgggcaa aacttattgc tgtgtcaacc atccttgtgc
     1681 ctgtcgtcag cattgcctcc tacactggat tggcgagcgg cctgacaatt tccgttcttg
     1741 aaatgccagc gggccatttt gcagaaggca gctcagtgat gctgggagga gaagaggtag
     1801 atggtgtagt caccatgtgg ggacggtatc tcacctgggc actttccacg cccatgattc
     1861 tcctcgctct gggtctcctg gccggaagca atgctacaaa gctcttcaca gctatcactt
     1921 tcgatatcgc tatgtgcgtg actggccttg ccgcggccct gactacctcc tcccacctca
     1981 tgagatggtt ctggtacgct atcagttgtg catgctttct ggtggtcttg tatatcctgc
     2041 tggtggagtg ggcacaggac gccaaagccg cgggaaccgc tgacatgttc aataccctga
     2101 agctgttgac agtagtgatg tggctggggt atccaattgt gtgggctctt ggagtcgagg
     2161 gtatcgcggt gttgcccgtt ggggtgacga gctggggata ttctttcctg gatatcgtgg
     2221 caaagtacat tttcgcattc ttgctcctga actatctgac gtcaaacgaa tctgtcgtgt
     2281 ccggcagcat tttggatgtt ccatctgctt ctgggacccc ggctgatgat gcggccgcag
     2341 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2401 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2461 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2521 tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     2581 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2641 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2701 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2761 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     2821 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     2881 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2941 tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3001 tggagttcgt gaccgccgcc gggggtacca aggtgtacga ccccgagcag aggaagagga
     3061 tgatcaccgg cccccagtgg tgggccaggt gcaagcagat gaacgtgctg gacagcttca
     3121 tcaactacta cgacagcgag aagcacgccg agaacgccgt gatcttcctg cacggcaacg
     3181 ccactagcag ctacctgtgg aggcacgtgg tgccccacat cgagcccgtg gccaggtgca
     3241 tcatccccga tctgatcggc atgggcaaga gcggcaagag cggcaacggc agctacaggc
     3301 tgctggacca ctacaagtac ctgaccgcct ggttcgagct tctgaacctg cccaagaaga
     3361 tcatcttcgt gggccacgac tggggcgccg ccctggcctt ccactacgcc tacgagcacc
     3421 aggacaggat caaggccatc gtgcacatgg agagcgtggt ggacgtgatc gagagctggg
     3481 acgagtggcc agacatcgag gaggacatcg ccctgatcaa gagcgaggag ggcgagaaga
     3541 tggtgctgga gaacaacttc ttcgtggaga ccgtgctgcc cagcaagatc atgagaaagc
     3601 tggagcccga ggagttcgcc gcctacctgg agcccttcaa ggagaagggc gaggtgagaa
     3661 gacccaccct gagctggccc agagagatcc ccctggtgaa gggcggcaag cccgacgtgg
     3721 tgcagatcgt gagaaactac aacgcctacc tgagagccag cgacgacctg cccaagctgt
     3781 tcatcgaggg cgaccccggc ttcttcagca acgccatcgt ggagggcgcc aagaagttcc
     3841 ccaacaccga gttcgtgaag gtgaagggcc tgcacttcct ccaggaggac gcccccgacg
     3901 agatgggcaa gtacatcaag agcttcgtgg agagagtgct gaagaacgag cagttctgct
     3961 acgagaacga ggtgtaagaa ttcgatatca agcttatcga taatcaacct ctggattaca
     4021 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     4081 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     4141 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     4201 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     4261 cctgtcagct cctttccgga actttcgctt tccccctccc tattgccacg gcggaactca
     4321 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     4381 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     4441 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     4501 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     4561 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     4621 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     4681 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     4741 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     4801 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     4861 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     4921 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     4981 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     5041 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     5101 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     5161 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     5221 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     5281 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5341 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5401 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     5461 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5521 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5581 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5641 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     5701 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     5761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5821 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5881 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5941 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6001 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6421 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6721 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7861 cttttgctca catgt
//
