LOCUS       sequence-199355-        7398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Ef1a-driven, cre-dependent, chimeric opsin variant
            C1V1 (E122T/E162T) for fast and potent optical excitation at 
            red-shifted wavelengths..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7398)
  AUTHORS   Yizhar O, Fenno LE, Prigge M, Schneider F, Davidson TJ, O'Shea DJ, 
            Sohal VS, Goshen I, Finkelstein J, Paz JT, Stehfest K, Fudim R, 
            Ramakrishnan C, Huguenard JR, Hegemann P, Deisseroth K
  TITLE     Neocortical excitation/inhibition balance in information processing 
            and social dysfunction.
  JOURNAL   Nature. 2011 Jul 27;477(7363):171-8. doi: 10.1038/nature10360.
  PUBMED    21796121
REFERENCE   2  (bases 1 to 7398)
  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"; journalName: 
            "Nature"; date: "2011-07-27- 27"; volume: "477"; issue: "7363"; 
            pages: "171-8"
COMMENT     Sequence Label: pAAV-Ef1a-DIO C1V1 (t/t)-TS-mCherry
FEATURES             Location/Qualifiers
     source          1..7398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        226..1404
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          457..1395
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1352..1372
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1435..1468)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1519..1552)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1564..2271)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1842..1861)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2111..2129
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    3383..3416
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3467..3500
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3525..4113
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3578..3598)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3996..4007)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4145..4621
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4344..4363)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4661..4801
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4661..4790
                     /label=AAV2 ITR
     rep_origin      4876..5331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4963..4982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5173..5194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5348..5367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5467..5489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5527..5545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5613..5717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5718..5786,5787..6578)
                     /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(5936..5955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6749..7337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7238..7257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgg gcgnnnnnna cctttggtcg
       61 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      121 ttcctgcggc cgcacgcgta agctttgcaa agatggataa agttttaaac agagaggaat
      181 ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt
      241 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
      301 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
      361 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
      421 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
      481 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
      541 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
      601 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
      661 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
      721 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
      781 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
      841 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
      901 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
      961 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     1021 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     1081 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     1141 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     1201 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     1261 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     1321 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     1381 tttcttccat ttcaggtgtc gtgaggtacc ggatcctcta gagtcgactc cggaataact
     1441 tcgtatagga tactttatac gaagttatgc agaatggtag ctggattgta gctgctatta
     1501 gcaatatgaa acctcttaat aacttcgtat agcatacatt atacgaagtt atggcgcgcc
     1561 ctattacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta
     1621 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt
     1681 gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc
     1741 gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc
     1801 ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg
     1861 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
     1921 cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc
     1981 ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt
     2041 cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag
     2101 ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
     2161 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa
     2221 gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctca ccacgttgat
     2281 gtcgatctgg tccaggggga tgtactcgcc ctcgctggtg atcctgctct tggcggccgc
     2341 gtcctcctct tcagccacca gtgtctcaac ctccatctct tggccggcga tcgtaatttt
     2401 ctgcttcttt ctgatgtcgc catacaacag gatgtgctcg tggattttga cgcgcaggta
     2461 attccccagc acaccccaca tgttctttgc aatcaggtcg aggatggagt gcccaatggc
     2521 agatccataa gggctgatat gaccgaatcc ttccgtgccc agcagaaaca ggactggaaa
     2581 cattccccaa gccacaaaga atgtccaggc catcactcgg accagttctc gacaaatccc
     2641 ttttggaact gtgtggaatg cttcgatata aactttagca gcatgaaaat aggtatacat
     2701 accatagctc aatgagatga ggaagaacag gatcttcgtc caccccgtac acatggcgct
     2761 ggttgcgccc cacacgatgc atcccacatc agaaaccaac aggcccatag tccttttgct
     2821 ataatcgtcc ttcaggcctg tgaggttgct cagatggatc agaaggaccg ggcaggtgag
     2881 cagccaggtc gcgtaacgaa gccacacggt cttgttgccg ttggatgagt agatcaccgc
     2941 aggttcgtca aactcatgga aatactcgat gatgaacttg atcatctcga tcgtggccac
     3001 gtaaatggtc tcccagccgc aagtagactt ccaggtctgg tagccgtaga acatcaggca
     3061 gagcgctgaa agcgcaaaag taatccactg cagaatgttg gcagccaact tctcggcatt
     3121 tgttccgttg gatttaagcc aagccaagca gaagcactgg ccgttgttcg ggatgcaaat
     3181 aacagaacca ttgttctcaa gagtgtatga ggtttggaag agcatgcgct cgtgggcacg
     3241 gtggaaaacg tagtcggggc catcctgagt cgcgaccggc accgtcgcgt cactgccagt
     3301 cgaggctcct gcgctgccgg ccgccagcgc cactgccagc gctagggcaa gaagccatgg
     3361 cctccgcgac atggtggcta gcataacttc gtataaagta tcctatacga agttatttgc
     3421 cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa
     3481 tgtatgctat acgaagttat gaattcgata tcaagcttat cgataatcaa cctctggatt
     3541 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3601 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3661 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3721 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3781 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3841 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3901 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     3961 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4021 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4081 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     4141 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     4201 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     4261 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     4321 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     4381 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     4441 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     4501 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     4561 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     4621 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     4681 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     4741 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     4801 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4861 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4921 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     4981 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5041 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     5101 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5161 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     5221 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     5281 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     5341 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5401 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     5461 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     5521 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5581 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5641 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5701 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5761 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5821 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5881 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5941 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6001 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6061 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6121 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6181 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6241 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6301 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6361 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6421 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6481 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6541 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6601 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6661 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6721 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6841 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6901 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6961 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7021 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7081 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7141 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7201 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7261 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7321 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7381 ggccttttgc tcacatgt
//
