LOCUS       sequence-196845-        7412 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 7412)
  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 7412)
  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-EYFP
FEATURES             Location/Qualifiers
     source          1..7412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   81..210
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        311..1489
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          542..1480
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1437..1457
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1520..1553)
                     /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(1604..1637)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1649..2362,2363..2365))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1694..1715)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2043..2062
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2302..2323
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    3477..3510
                     /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    3561..3594
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3619..4207
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3672..3692)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4090..4101)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4239..4715
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4438..4457)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4755..4895
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4755..4884
                     /label=AAV2 ITR
     rep_origin      4970..5425
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5057..5076)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5267..5288
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5442..5461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5561..5583
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5621..5639)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5707..5811
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5812..5880,5881..6672)
                     /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(6030..6049)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6843..7412,1..19)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7332..7351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
       61 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
      121 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      181 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtaagctt tgcaaagatg
      241 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg
      301 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
      361 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
      421 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
      481 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
      541 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
      601 gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
      661 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
      721 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
      781 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
      841 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
      901 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
      961 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     1021 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     1081 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     1141 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     1201 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     1261 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     1321 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     1381 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     1441 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc
     1501 ctctagagtc gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat
     1561 ggtagctgga ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat
     1621 acattatacg aagttatggc gcgccctatt acttgtacag ctcgtccatg ccgagagtga
     1681 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     1741 tgctcagggc ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     1801 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     1861 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     1921 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     1981 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     2041 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     2101 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact
     2161 gcaggccgta gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     2221 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     2281 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     2341 tgaacagctc ctcgcccttg ctcaccacgt tgatgtcgat ctggtccagg gggatgtact
     2401 cgccctcgct ggtgatcctg ctcttggcgg ccgcgtcctc ctcttcagcc accagtgtct
     2461 caacctccat ctcttggccg gcgatcgtaa ttttctgctt ctttctgatg tcgccataca
     2521 acaggatgtg ctcgtggatt ttgacgcgca ggtaattccc cagcacaccc cacatgttct
     2581 ttgcaatcag gtcgaggatg gagtgcccaa tggcagatcc ataagggctg atatgaccga
     2641 atccttccgt gcccagcaga aacaggactg gaaacattcc ccaagccaca aagaatgtcc
     2701 aggccatcac tcggaccagt tctcgacaaa tcccttttgg aactgtgtgg aatgcttcga
     2761 tataaacttt agcagcatga aaataggtat acataccata gctcaatgag atgaggaaga
     2821 acaggatctt cgtccacccc gtacacatgg cgctggttgc gccccacacg atgcatccca
     2881 catcagaaac caacaggccc atagtccttt tgctataatc gtccttcagg cctgtgaggt
     2941 tgctcagatg gatcagaagg accgggcagg tgagcagcca ggtcgcgtaa cgaagccaca
     3001 cggtcttgtt gccgttggat gagtagatca ccgcaggttc gtcaaactca tggaaatact
     3061 cgatgatgaa cttgatcatc tcgatcgtgg ccacgtaaat ggtctcccag ccgcaagtag
     3121 acttccaggt ctggtagccg tagaacatca ggcagagcgc tgaaagcgca aaagtaatcc
     3181 actgcagaat gttggcagcc aacttctcgg catttgttcc gttggattta agccaagcca
     3241 agcagaagca ctggccgttg ttcgggatgc aaataacaga accattgttc tcaagagtgt
     3301 atgaggtttg gaagagcatg cgctcgtggg cacggtggaa aacgtagtcg gggccatcct
     3361 gagtcgcgac cggcaccgtc gcgtcactgc cagtcgaggc tcctgcgctg ccggccgcca
     3421 gcgccactgc cagcgctagg gcaagaagcc atggcctccg cgacatggtg gctagcataa
     3481 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
     3541 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc
     3601 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3661 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3721 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3781 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3841 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3901 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3961 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     4021 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     4081 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     4141 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     4201 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     4261 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     4321 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     4381 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     4441 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     4501 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     4561 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     4621 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     4681 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     4741 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4801 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4861 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     4921 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     4981 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5041 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5101 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5161 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     5221 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5281 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     5341 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5401 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     5461 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5521 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5581 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5641 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5701 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5761 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5821 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5881 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5941 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6001 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6061 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6121 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6181 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6241 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6301 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6361 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6421 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6481 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6541 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6601 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6661 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6721 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6781 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6841 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6901 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6961 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7021 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7081 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7141 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7201 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7261 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7321 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7381 cttgagcgtc gatttttgtg atgctcgtca gg
//
