LOCUS       sequence-212375-        6886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with hSynapsin promoter, Cre-OFF mCAR (for efficient 
            CAV-2 infection) and Cre-ON ChETA (for optogenetic activation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6886)
  AUTHORS   Li SJ, Vaughan A, Sturgill JF, Kepecs A
  TITLE     A Viral Receptor Complementation Strategy to Overcome CAV-2 Tropism 
            for Efficient Retrograde Targeting of Neurons.
  JOURNAL   Neuron. 2018 Jun 6;98(5):905-917.e5. doi: 
            10.1016/j.neuron.2018.05.028.
  PUBMED    29879392
REFERENCE   2  (bases 1 to 6886)
  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: 
            "10.1016/j.neuron.2018.05"; journalName: "Neuron"; date: "2018-06-6-
            6"; volume: "98"; issue: "5"; pages: "905-917.e5"
COMMENT     Sequence Label: pAAV-hSyn-DIO {mCAR}off-{ChETA}on-WPRE
FEATURES             Location/Qualifiers
     source          1..6886
                     /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        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /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(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      783..792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1845..1874
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1889..1915)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(1889..1915)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1934..2860)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYATWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2857..2866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2871..2904
                     /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    2955..2988
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3013..3601
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3066..3086)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3484..3495)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3633..4109
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3832..3851)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4149..4289
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4149..4278
                     /label=AAV2 ITR
     rep_origin      4364..4819
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4451..4470)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4661..4682
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4836..4855)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4955..4977
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5015..5033)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5101..5205
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5206..5274,5275..6066)
                     /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(5424..5443)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6237..6825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6726..6745
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccaccat ggctcgccta ctgtgcttcg tgctcttgtg cgggatcgcg gatttcacca
      841 gtggtttgag catcactaca cccgaacaga ggatcgaaaa agccaaaggg gaaactgcgt
      901 atctaccatg caagtttact ctcagtcccg aagaccaggg accactggac attgaatggc
      961 tgatatcccc gtctgataac cagatagtgg atcaagtgat cattttgtat tctggagaca
     1021 aaatttatga taactactat ccggatctga aaggacgggt acattttacg agtaacgatg
     1081 tcaagtctgg cgacgcatct ataaatgtga ccaacctgca gctgtcggac attggcactt
     1141 accagtgcaa agtgaagaaa gcccctgggg ttgcaaataa gaaattcctg ctgaccgttc
     1201 ttgttaagcc ttcaggtaca agatgcttcg tggatggatc ggaagagatt ggaaatgact
     1261 tcaagctaaa atgtgaaccc aaggaaggct cccttccact acagtttgaa tggcagaaac
     1321 tgtcggactc ccagacaatg cctacgccat ggctggcaga aatgacgtca ccagttatat
     1381 ctgtgaagaa cgccagttct gagtattctg ggacatacag ctgcacggtt caaaacagag
     1441 tgggctctga ccagtgtatg ctgcgactag acgttgtccc accctccaac cgagccggaa
     1501 cgatcgcggg cgccgtcata gggacgctgc tggcccttgt gctcatcggg gccatcctct
     1561 tctgctgtca caggaaacgc agagaagaga agtacgagaa ggaagttcat catgatatca
     1621 gggaagatgt gcctcctcca aagagtcgga catccactgc caggagctat attggcagca
     1681 accattcctc cctgggatcc atgtccccct ctaacatgga ggggtattcc aagacgcagt
     1741 ataaccaagt ccccagtgaa gactttgaac gtgcgcctca gagcccgact ctggcacccg
     1801 ctaagttcaa gtacgcttac aagaccgatg gcattacagt ggtagagcag aagctgatca
     1861 gcgaggagga cctgtaatta attaattagg cgtagtcggg cacgtcgtag gggtacatgg
     1921 tggcggccgc tggcacggct ccggcctcgg cttcgtcttc gacgagagtc tcgacctcga
     1981 tctccgttcc gccgatgttc agtttggtgg ttttgcggat atctccgtga atcaatatgt
     2041 gctcgtggat caggacgcgc aggtagtgtc ccaacaaccc ccaacaattt ttactcatca
     2101 gatcaataat cgtgtgacct acggtggagc catagacgct caggacgcca aaaccttcgg
     2161 gccccaaaat gaagagaatt gggaacatac cccagctcac gaaaaacagc catgccatgc
     2221 cggtcacgac ctggcggcac cgaccctttg gcacagtatg ataaccctcg atatatgctt
     2281 tggcggcgtg aaaaaatgtg ttcgcgccat agcacaatcc aagacaaaag aagatgactt
     2341 taacatagcc ggttgccatg gcgctggtag ccccccacac gatagtcccg atgtctgaga
     2401 caaggagtcc catggttctc ctgctgtagt cgttgctcag gccggtgagg ttgctcaggc
     2461 ggataaggat gacaggacaa gtgagcagcc aggttgcata gcgcagccac tgcacccggt
     2521 gtcctgtggc aaggtagagc atagagggat tcttaaactc aaaaaagaac tcgagaatca
     2581 ccttaaccat ttcaatggcg cacacataga tctcctccca gccgcatgta gatttccagg
     2641 tttggtaggc atagaacatc agcagcaaaa tgctgaatcc tgctgcaagc cactgcagga
     2701 catttgacgc ggtctgagcg ccgttcgtgc cgcgagattc aatccatccg gcacagtaac
     2761 attgatcctc agggaccagg acggacccgt tcaccaccac aggattagta acgaacaaaa
     2821 gttcgcgtcc gacggcagac aaagcgccgc catagtccat ggtggctagc ataacttcgt
     2881 ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg ttattcttta
     2941 gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga attcgatatc
     3001 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3061 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3121 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3181 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3241 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3301 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3361 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3421 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3481 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3541 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3601 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     3661 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     3721 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     3781 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     3841 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     3901 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     3961 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     4021 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     4081 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     4141 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4201 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4261 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4321 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4381 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4441 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4501 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4561 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4621 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4681 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4741 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4801 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4861 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4921 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4981 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5041 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5101 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5161 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5221 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5281 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5341 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5401 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5461 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5521 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5581 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5641 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5701 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5761 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5821 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5881 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5941 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6001 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6061 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6121 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6181 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6241 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6301 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6361 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6421 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6481 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6541 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6601 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6661 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6721 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6781 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6841 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
