LOCUS       sequence-209264-        7287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with hSynapsin promoter, Cre-OFF mCAR (for efficient 
            CAV-2 infection), Cre-ON DTR-EGFP (diphtheria toxin receptor for 
            cell ablation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7287)
  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 7287)
  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-{DTR-GFP}on-WPRE
FEATURES             Location/Qualifiers
     source          1..7287
                     /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(join(1898..2611,2612..2614,2615..2617))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1901..2611,2612..2614,2615..2617))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1943..1964)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2292..2311
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2551..2572
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2614..2623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3273..3306
                     /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    3357..3390
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3415..4003
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3468..3488)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3886..3897)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4035..4511
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4234..4253)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4551..4691
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4551..4680
                     /label=AAV2 ITR
     rep_origin      4766..5221
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4853..4872)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5063..5084
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5238..5257)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5357..5379
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5417..5435)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5503..5607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5608..5676,5677..6468)
                     /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(5826..5845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6639..7227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7128..7147
                     /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 cttgtacagc tcgtccatgc
     1921 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     1981 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     2041 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     2101 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     2161 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     2221 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     2281 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     2341 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     2401 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     2461 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     2521 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     2581 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatccgggt
     2641 gggaattagt catgcccaac ttcactttct cttcgttttc cacatcataa ccacctctcc
     2701 tatggtacct aaacatgaga agccccacga tgaccagcag acagacagag gacagcacca
     2761 cggccaccac agccaggata gttgtatggt cataggtata taagcgattt tccactggga
     2821 ggctcagccc atgacacctc tctccatggt aacctgggtg gcagatgcag gagggagccc
     2881 ggagctcctt cacatatttg cattctccgt ggatgcagaa gtccttgtat ttccgaagac
     2941 atgggtccct cttcttccct agtcccttgc ctttcttctt tcttttcccg tgctcctcct
     3001 tgcttggtgt ggccagtgct tgtggcttgg aggataaagt gactctcaaa aggtccagat
     3061 ctgcctcttg caagtcacgg actttccggt cccggccgcc tcctaggcgt agcagctggt
     3121 ccgtagatcc agtggaaggg tccgggttgc tggttccagc agctagccct ctccgaagct
     3181 gctccaggct ctcgccagtc accagtgccg aaagaactgc agccagaagg agcttcagca
     3241 ccaccgacgg cagcagcttc atggtgacta gcataacttc gtataaagta tcctatacga
     3301 agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa
     3361 cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat cgataatcaa
     3421 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3481 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3541 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3601 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3661 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3721 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3781 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     3841 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3901 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3961 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     4021 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     4081 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     4141 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     4201 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     4261 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     4321 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     4381 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     4441 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     4501 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     4561 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4621 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4681 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4741 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4921 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4981 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5101 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5161 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5221 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5281 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5341 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5401 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5461 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5761 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5821 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5881 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6121 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6181 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6241 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6301 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6361 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6421 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6481 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6781 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7261 gccttttgct ggccttttgc tcacatg
//
