LOCUS       sequence-194678-        7403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of ChR1-VChR1 Chimera fused to mCherry driven by Ef1a
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  TITLE     Deisseroth AAV plasmids
REFERENCE   2  (bases 1 to 7403)
  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"
COMMENT     Sequence Label: pAAV-Ef1a-DIO C1V1 (t/t)-TS-mCherry (PV2670)
FEATURES             Location/Qualifiers
     source          1..7403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     173..192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   334..463
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        564..1742
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          795..1733
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1690..1710
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1773..1806)
                     /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(1857..1890)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1902..2609)
                     /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(2180..2199)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2449..2467
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    3721..3754
                     /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    3805..3838
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3863..4451
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3916..3936)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4334..4345)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4483..4959
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4682..4701)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4999..5139
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4999..5128
                     /label=AAV2 ITR
     rep_origin      5214..5669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5301..5320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5511..5532
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5686..5705)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5805..5827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5865..5883)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5951..6055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6056..6124,6125..6916)
                     /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(6274..6293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7087..7403,1..272)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
       61 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      181 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      301 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
      361 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
      421 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtaag
      481 ctttgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct
      541 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
      601 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
      661 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      721 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      781 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
      841 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
      901 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
      961 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
     1021 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
     1081 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     1141 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     1201 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     1261 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     1321 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     1381 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     1441 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     1501 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     1561 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     1621 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     1681 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     1741 gaggtaccgg atcctctaga gtcgactccg gaataacttc gtataggata ctttatacga
     1801 agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttaataa
     1861 cttcgtatag catacattat acgaagttat ggcgcgccct attacttgta cagctcgtcc
     1921 atgccgccgg tggagtggcg gccctcggcg cgttcgtact gttccacgat ggtgtagtcc
     1981 tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt aggcgccggg cagctgcacg
     2041 ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc
     2101 ttcagcctct gcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggag
     2161 gaggcctccc agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg
     2221 ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg
     2281 gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag
     2341 gacagcttca agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac
     2401 atgaactgag gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc
     2461 agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac
     2521 tcgtggccgt tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg
     2581 gccatgttat cctcctcgcc cttgctcacc acgttgatgt cgatctggtc cagggggatg
     2641 tactcgccct cgctggtgat cctgctcttg gcggccgcgt cctcctcttc agccaccagt
     2701 gtctcaacct ccatctcttg gccggcgatc gtaattttct gcttctttct gatgtcgcca
     2761 tacaacagga tgtgctcgtg gattttgacg cgcaggtaat tccccagcac accccacatg
     2821 ttctttgcaa tcaggtcgag gatggagtgc ccaatggcag atccataagg gctgatatga
     2881 ccgaatcctt ccgtgcccag cagaaacagg actggaaaca ttccccaagc cacaaagaat
     2941 gtccaggcca tcactcggac cagttctcga caaatccctt ttggaactgt gtggaatgct
     3001 tcgatataaa ctttagcagc atgaaaatag gtatacatac catagctcaa tgagatgagg
     3061 aagaacagga tcttcgtcca ccccgtacac atggcgctgg ttgcgcccca cacgatgcat
     3121 cccacatcag aaaccaacag gcccatagtc cttttgctat aatcgtcctt caggcctgtg
     3181 aggttgctca gatggatcag aaggaccggg caggtgagca gccaggtcgc gtaacgaagc
     3241 cacacggtct tgttgccgtt ggatgagtag atcaccgcag gttcgtcaaa ctcatggaaa
     3301 tactcgatga tgaacttgat catctcgatc gtggccacgt aaatggtctc ccagccgcaa
     3361 gtagacttcc aggtctggta gccgtagaac atcaggcaga gcgctgaaag cgcaaaagta
     3421 atccactgca gaatgttggc agccaacttc tcggcatttg ttccgttgga tttaagccaa
     3481 gccaagcaga agcactggcc gttgttcggg atgcaaataa cagaaccatt gttctcaaga
     3541 gtgtatgagg tttggaagag catgcgctcg tgggcacggt ggaaaacgta gtcggggcca
     3601 tcctgagtcg cgaccggcac cgtcgcgtca ctgccagtcg aggctcctgc gctgccggcc
     3661 gccagcgcca ctgccagcgc tagggcaaga agccatggcc tccgcgacat ggtggctagc
     3721 ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg
     3781 ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga
     3841 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3901 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3961 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4021 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4081 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4141 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4201 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4261 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     4321 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4381 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4441 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     4501 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     4561 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4621 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4681 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4741 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4801 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4861 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4921 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4981 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     5041 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     5101 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     5161 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     5221 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5281 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5341 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5401 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     5461 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5521 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     5581 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5641 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5701 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5761 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5821 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5881 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5941 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6001 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6061 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6121 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6181 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6241 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6301 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6361 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6421 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6481 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6541 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6601 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6661 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6721 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6781 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6841 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6901 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6961 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7201 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7381 ggataaggcg cagcggtcgg gct
//
