LOCUS       sequence-43572-e        7413 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 7413)
  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 7413)
  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..7413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1450..1483)
                     /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(1534..1567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1579..2286)
                     /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(1857..1876)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2126..2144
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    3398..3431
                     /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    3482..3515
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3540..4128
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3593..3613)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4011..4022)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4160..4636
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4359..4378)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4676..4816
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4676..4805
                     /label=AAV2 ITR
     rep_origin      4891..5346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4978..4997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5188..5209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5363..5382)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5482..5504
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5542..5560)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5628..5732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5733..5801,5802..6593)
                     /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(5951..5970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6764..7352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7253..7272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccctatt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc
     1621 ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa
     1681 cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt
     1741 cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc
     1801 cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt
     1861 cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat
     1921 gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
     1981 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat
     2041 gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc
     2101 ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
     2161 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
     2221 catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt
     2281 gctcaccacg ttgatgtcga tctggtccag ggggatgtac tcgccctcgc tggtgatcct
     2341 gctcttggcg gccgcgtcct cctcttcagc caccagtgtc tcaacctcca tctcttggcc
     2401 ggcgatcgta attttctgct tctttctgat gtcgccatac aacaggatgt gctcgtggat
     2461 tttgacgcgc aggtaattcc ccagcacacc ccacatgttc tttgcaatca ggtcgaggat
     2521 ggagtgccca atggcagatc cataagggct gatatgaccg aatccttccg tgcccagcag
     2581 aaacaggact ggaaacattc cccaagccac aaagaatgtc caggccatca ctcggaccag
     2641 ttctcgacaa atcccttttg gaactgtgtg gaatgcttcg atataaactt tagcagcatg
     2701 aaaataggta tacataccat agctcaatga gatgaggaag aacaggatct tcgtccaccc
     2761 cgtacacatg gcgctggttg cgccccacac gatgcatccc acatcagaaa ccaacaggcc
     2821 catagtcctt ttgctataat cgtccttcag gcctgtgagg ttgctcagat ggatcagaag
     2881 gaccgggcag gtgagcagcc aggtcgcgta acgaagccac acggtcttgt tgccgttgga
     2941 tgagtagatc accgcaggtt cgtcaaactc atggaaatac tcgatgatga acttgatcat
     3001 ctcgatcgtg gccacgtaaa tggtctccca gccgcaagta gacttccagg tctggtagcc
     3061 gtagaacatc aggcagagcg ctgaaagcgc aaaagtaatc cactgcagaa tgttggcagc
     3121 caacttctcg gcatttgttc cgttggattt aagccaagcc aagcagaagc actggccgtt
     3181 gttcgggatg caaataacag aaccattgtt ctcaagagtg tatgaggttt ggaagagcat
     3241 gcgctcgtgg gcacggtgga aaacgtagtc ggggccatcc tgagtcgcga ccggcaccgt
     3301 cgcgtcactg ccagtcgagg ctcctgcgct gccggccgcc agcgccactg ccagcgctag
     3361 ggcaagaagc catggcctcc gcgacatggt ggctagcata acttcgtata aagtatccta
     3421 tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     3481 aataacttcg tataatgtat gctatacgaa gttatgaatt cgatatcaag cttatcgata
     3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3661 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3721 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3781 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3841 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3901 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3961 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4021 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4081 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     4141 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     4201 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4261 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4321 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     4381 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     4441 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     4501 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     4561 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     4621 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     4681 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4741 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4801 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4861 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4921 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4981 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5041 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5101 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5161 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5221 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5281 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5341 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5401 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5461 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5521 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5581 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5701 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5761 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5821 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5881 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5941 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6001 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6061 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6121 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6181 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6241 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6301 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6361 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6421 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6481 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6541 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6601 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6661 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6721 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6781 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6841 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6901 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6961 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7021 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7081 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7141 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7201 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7261 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7321 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7381 tcctggcctt ttgctggcct tttgctcaca tgt
//
