LOCUS       sequence-190877-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome with Cre-dependent expression of the tTA from the 
            hSyn1 promoter. Contains a positive feedback loop for amplifed 
            expression of the tTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5729)
  AUTHORS   Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, 
            Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V
  TITLE     Engineered AAVs for efficient noninvasive gene delivery to the 
            central and peripheral nervous systems.
  JOURNAL   Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 
            2017 Jun 26.
  PUBMED    28671695
REFERENCE   2  (bases 1 to 5729)
  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: "10.1038/nn.4593"; 
            journalName: "Nat Neurosci"; date: "2017-08"; volume: "20"; issue: 
            "8"; pages: "1172-1179"
COMMENT     Sequence Label: pAAV-ihSyn1-DIO-tTA
FEATURES             Location/Qualifiers
     source          1..5729
                     /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    662..680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    698..716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(829..862)
                     /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(913..946)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(964..1074)
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     protein_bind    1714..1747
                     /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    1798..1831
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1856..2444
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1909..1929)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2327..2338)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2476..2952
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2675..2694)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2992..3132
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2992..3121
                     /label=AAV2 ITR
     rep_origin      3207..3662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3294..3313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3504..3525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3679..3698)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3798..3820
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3858..3876)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3944..4048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4049..4117,4118..4909)
                     /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(4267..4286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5080..5668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5569..5588
                     /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 agtcgagaac agatctctaa aacaggtaag tcccattaat
      661 ctccctatca gtgatagaga aggtctgaag agtttactcc ctatcagtga tagagattaa
      721 ttcctctact aaccttgttc atcttttctt tttttttcta caggtcctgg gtgattaaca
      781 gcttaagggc ccggatccgg taccggatcc tctagagtcg actccggaat aacttcgtat
      841 aggatacttt atacgaagtt atgcagaatg gtagctggat tgtagctgct attagcaata
      901 tgaaacctct taataacttc gtatagcata cattatacga agttatggcg cgccttaccc
      961 ggggagcatg tcaaggtcaa aatcgtcaag agcgtcagca ggcagcatat caaggtcaaa
     1021 gtcgtcaagg gcatcggctg ggagcatgtc taagtcaaaa tcgtcaaggg cgtcggccgg
     1081 cccgccgctt tcgcacttta gctgtttctc caggccacat atgattagtt ccaggccgaa
     1141 aaggaaggca ggttcggctc cctgccggtc gaacagctca attgcttgtc tcagaagtgg
     1201 gggcatagaa tcggtggtag gtgtctctct ttcctctttt gctacttgat gctcctgttc
     1261 ctccaatacg cagcccagtg taaagtggcc cacggcggac agagcgtaca gtgcgttctc
     1321 cagggagaag ccttgctgac acaggaacgc gagctgattt tccagggttt cgtactgttt
     1381 ctctgttggg cgggtgccga gatgcacttt agccccgtcg cgatgtgaga ggagagcaca
     1441 gcggaatgac ttggcgttgt tccgcagaaa gtcttgccat gactcgcctt ccagggggca
     1501 gaagtgggta tgatgcctgt ccagcatctc gatggccagg gcatcgagca gggcccgctt
     1561 gttcttcacg tgccagtaca gggtaggctg ctcaactccc agcttttgag cgagtttcct
     1621 tgtcgtcagg ccttcgatac cgacttcatt gagtaattcc agagcgccgt ttatgacttt
     1681 agatttatcc agccgactca tggtggcgct agcataactt cgtataaagt atcctatacg
     1741 aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata
     1801 acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca
     1861 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     1921 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     1981 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2041 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2101 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2161 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2221 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
     2281 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2341 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2401 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
     2461 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     2521 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     2581 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     2641 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     2701 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     2761 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     2821 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     2881 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     2941 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     3001 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3061 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3121 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3181 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3241 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3301 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3361 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3421 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3481 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3541 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3601 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3661 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3721 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3781 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3841 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3901 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3961 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4021 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4081 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4141 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4201 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4261 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4321 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4381 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4441 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4501 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4561 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4621 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4681 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4741 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4801 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4861 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4921 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4981 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5041 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5101 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5161 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5221 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5281 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5341 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5401 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5461 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5521 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5581 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5641 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5701 ggccttttgc tggccttttg ctcacatgt
//
