LOCUS       sequence-209248-        6897 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 6897)
  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 6897)
  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..6897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..142
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   13..142
                     /label=AAV2 ITR
     promoter        187..634
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(672..705)
                     /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(756..789)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      795..804
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1857..1886
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1901..1927)
                     /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(1901..1927)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1946..2872)
                     /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      2869..2878
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2883..2916
                     /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    2967..3000
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3025..3613
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3078..3098)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3496..3507)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3645..4121
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3844..3863)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4161..4301
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4161..4290
                     /label=AAV2 ITR
     rep_origin      4376..4831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4463..4482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4673..4694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4848..4867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4967..4989
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5027..5045)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5113..5217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5218..5286,5287..6078)
                     /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(5436..5455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6249..6837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6738..6757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca gagggccctg
      181 cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac
      241 gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct
      301 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc
      361 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga
      421 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg
      481 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca
      541 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg
      601 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga tcctctagag
      661 tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg
      721 gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata
      781 cgaagttatg gcgcgccacc atggctcgcc tactgtgctt cgtgctcttg tgcgggatcg
      841 cggatttcac cagtggtttg agcatcacta cacccgaaca gaggatcgaa aaagccaaag
      901 gggaaactgc gtatctacca tgcaagttta ctctcagtcc cgaagaccag ggaccactgg
      961 acattgaatg gctgatatcc ccgtctgata accagatagt ggatcaagtg atcattttgt
     1021 attctggaga caaaatttat gataactact atccggatct gaaaggacgg gtacatttta
     1081 cgagtaacga tgtcaagtct ggcgacgcat ctataaatgt gaccaacctg cagctgtcgg
     1141 acattggcac ttaccagtgc aaagtgaaga aagcccctgg ggttgcaaat aagaaattcc
     1201 tgctgaccgt tcttgttaag ccttcaggta caagatgctt cgtggatgga tcggaagaga
     1261 ttggaaatga cttcaagcta aaatgtgaac ccaaggaagg ctcccttcca ctacagtttg
     1321 aatggcagaa actgtcggac tcccagacaa tgcctacgcc atggctggca gaaatgacgt
     1381 caccagttat atctgtgaag aacgccagtt ctgagtattc tgggacatac agctgcacgg
     1441 ttcaaaacag agtgggctct gaccagtgta tgctgcgact agacgttgtc ccaccctcca
     1501 accgagccgg aacgatcgcg ggcgccgtca tagggacgct gctggccctt gtgctcatcg
     1561 gggccatcct cttctgctgt cacaggaaac gcagagaaga gaagtacgag aaggaagttc
     1621 atcatgatat cagggaagat gtgcctcctc caaagagtcg gacatccact gccaggagct
     1681 atattggcag caaccattcc tccctgggat ccatgtcccc ctctaacatg gaggggtatt
     1741 ccaagacgca gtataaccaa gtccccagtg aagactttga acgtgcgcct cagagcccga
     1801 ctctggcacc cgctaagttc aagtacgctt acaagaccga tggcattaca gtggtagagc
     1861 agaagctgat cagcgaggag gacctgtaat taattaatta ggcgtagtcg ggcacgtcgt
     1921 aggggtacat ggtggcggcc gctggcacgg ctccggcctc ggcttcgtct tcgacgagag
     1981 tctcgacctc gatctccgtt ccgccgatgt tcagtttggt ggttttgcgg atatctccgt
     2041 gaatcaatat gtgctcgtgg atcaggacgc gcaggtagtg tcccaacaac ccccaacaat
     2101 ttttactcat cagatcaata atcgtgtgac ctacggtgga gccatagacg ctcaggacgc
     2161 caaaaccttc gggccccaaa atgaagagaa ttgggaacat accccagctc acgaaaaaca
     2221 gccatgccat gccggtcacg acctggcggc accgaccctt tggcacagta tgataaccct
     2281 cgatatatgc tttggcggcg tgaaaaaatg tgttcgcgcc atagcacaat ccaagacaaa
     2341 agaagatgac tttaacatag ccggttgcca tggcgctggt agccccccac acgatagtcc
     2401 cgatgtctga gacaaggagt cccatggttc tcctgctgta gtcgttgctc aggccggtga
     2461 ggttgctcag gcggataagg atgacaggac aagtgagcag ccaggttgca tagcgcagcc
     2521 actgcacccg gtgtcctgtg gcaaggtaga gcatagaggg attcttaaac tcaaaaaaga
     2581 actcgagaat caccttaacc atttcaatgg cgcacacata gatctcctcc cagccgcatg
     2641 tagatttcca ggtttggtag gcatagaaca tcagcagcaa aatgctgaat cctgctgcaa
     2701 gccactgcag gacatttgac gcggtctgag cgccgttcgt gccgcgagat tcaatccatc
     2761 cggcacagta acattgatcc tcagggacca ggacggaccc gttcaccacc acaggattag
     2821 taacgaacaa aagttcgcgt ccgacggcag acaaagcgcc gccatagtcc atggtggcta
     2881 gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac
     2941 tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat
     3001 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3061 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3121 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3181 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3241 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3301 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3361 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3421 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     3481 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3541 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3601 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     3661 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     3721 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     3781 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     3841 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     3901 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     3961 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     4021 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     4081 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     4141 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4201 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4261 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4321 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4381 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4441 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4501 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4561 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4621 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4681 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     4741 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4801 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4861 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4921 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4981 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5041 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5101 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5161 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5221 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5281 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5341 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5401 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5461 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5521 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5581 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5641 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5701 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5761 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5821 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5881 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5941 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6001 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6061 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6121 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6361 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6661 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatg
//
