LOCUS       sequence-354867-        6616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Flp-dependent transgene expression of 
            TeTLc-P2A-mCherry in neurons under the control of the hSyn1 promoter
            (Kugler et al 2003).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  AUTHORS   Pouchelon G, Vergara J, McMahon J, Gorissen BL, Lin JD, 
            Vormstein-Schneider D, Niehaus JL, Burbridge TJ, Wester JC, Sherer 
            M, Fernandez-Otero M, Allaway KC, Pelkey K, Chittajallu R, McBain 
            CJ, Fan M, Nasse JS, Wildenberg GA, Fishell G, Dimidschstein J
  TITLE     A versatile viral toolkit for functional discovery in the nervous 
            system.
  JOURNAL   Cell Rep Methods. 2022 May 26;2(6):100225. doi: 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20.
  PUBMED    35784651
REFERENCE   2  (bases 1 to 6616)
  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"; journalName: "Cell Rep 
            Methods."; date: "2022-05-26"; volume: "2(6):100225. doi"; pages: " 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20"
COMMENT     Sequence Label: pAAV-VTKS3-TeTLc-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      168..623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(255..274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     465..486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(640..659)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     759..781
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(819..837)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        905..1009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1010..1078,1079..1870)
                     /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(1228..1247)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2041..2629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2530..2549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2691..2820
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2848..3295
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    3319..3352
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(3459..4166)
                     /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(3737..3756)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4006..4024
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(4167..4223)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    complement(5621..5654)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    5747..6335
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5800..5820)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6218..6229)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6343..6567
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   join(6569..6616,1..93)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(6569..6616,1..82)
                     /label=AAV2 ITR
ORIGIN
        1 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
       61 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
      121 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
      181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
      241 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
      301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
      361 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
      421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
      481 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
      541 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
      601 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
      661 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      721 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      781 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
      841 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      901 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      961 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1021 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1081 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1141 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1201 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1261 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1321 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1381 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1441 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1501 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1561 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1621 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1681 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1741 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1801 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1861 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1921 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1981 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2161 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2281 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2341 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2401 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2461 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2521 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2581 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2641 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     2701 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
     2761 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     2821 gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg
     2881 gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat
     2941 tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc
     3001 gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg
     3061 ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc
     3121 ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc
     3181 gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc
     3241 gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcaga
     3301 tatcgtttaa actccggaga agttcctatt ctctagaaag tataggaact tcgcagaatg
     3361 gtagctggat tgtagctgct attagcaata tgaaacctct tagaagttcc tattcttcaa
     3421 aaggtatagg aacttcaacg cgccctagaa ttcgcgcctt acttgtacag ctcgtccatg
     3481 ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg
     3541 ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc
     3601 ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc
     3661 agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag
     3721 gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg
     3781 cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc
     3841 accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac
     3901 agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg
     3961 aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc
     4021 ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg
     4081 tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc
     4141 atgttatcct cctcgccctt gctcacggga ccggggtttt cttccacgtc tcctgcttgc
     4201 tttaacagag agaagttcgt ggctccgctt ccagcggtac ggttgtacag gttttcacgg
     4261 atgttggtcg gcgggatgat ttttttgcac agaccgatca gtttggaaac cagaccggaa
     4321 ccgtcgacgt tcctaaaggc gttagtgttc accctcatgt tctggccctt atactcggac
     4381 ttcagatcct tggactctat gttaaagccc tcagtatcgt tatatatagt atcatccagc
     4441 aggttaggta tcttcacagg atcgtggttc atggaaaaat agcttaagcg agtctttatg
     4501 ttaaacttct tgcccagctc tatctcagta aagccataca ttatggagtt atacagtatc
     4561 tgaaacttat cctcgttcac tatatactgg ccgttggaat ccttatcaaa ctgatacttc
     4621 tgctgataga tctgtttgta ggagtcgatg tcgatgttcg ggtcgttgca ggaggtaacc
     4681 tgggacagtt tgttagcgat agctttgtag tcgttcaggg ttttttcgta caggtcgttt
     4741 ttgatatcga tggagatcag gttagcgtcc tgaccaccga aggtgaagag ctcttcagcg
     4801 gagatcgggt aggtgtgctg catgtagatt tcctgtttgg acgggatgat ttcgtgggag
     4861 gaaacctgca tgccgtacag accgtgcaga acgtggatca gttcgtgcat cagcagcaga
     4921 gccgggtcct ggaagtattt ggatttaccg atggtcaggg aggtgatgtt ttcgataacg
     4981 ttgtcgaagg tcggtacgta ttccgggcag aaagccatct gcatgatgga accgaaaccg
     5041 tcacggcacg ggaagtagtt tttgttgtca acacgcagaa cgataccacg aacttcgttt
     5101 ttgttcagaa ccggacccgg accgaagatg atcaggttgg tcagcatagc ggatttggtg
     5161 gtagcaccgg acggatcctg ttccagcagg ttgaaggaaa cggagttgga gttggtgtcg
     5221 aatttgtcca gcagggagta ggagttaccc aggtacggga tagcgttgat gattttgtcc
     5281 agcagagctt caccagcaac gttgtttttg atacggttga acagtttaac catggtctgc
     5341 aggaaacggt ctttgtcgga gtcggtacgc aggtagttcg ggtcgtagta ttcggaagca
     5401 ccttcgatca gggaggacgg cgggttgaag tcttccggtt tggtaccaaa ttcgtaacgt
     5461 tccggaacga tccagatacg gtcggtgatt ttgaaggctt tgtagtagat gtctagacct
     5521 ttgcagtacg gcggttccat catgatgatg gtgtcgttgt taaccgggtc ggagtaacgg
     5581 aagttgttga tggtgatcgg catggtggcg gctagctagc gaagttccta tactttctag
     5641 agaataggaa cttcttgcct taacccagaa attatcactg ttattcttta gaatggtgca
     5701 aagagaagtt cctatacctt ttgaagaata ggaacttcgg cgcgccaatc aacctctgga
     5761 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5821 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5881 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5941 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6001 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6061 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6121 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     6181 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6241 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6301 gacgagtcgg atctcccttt gggccgcctc cccgcctgga tcctgtgcct tctagttgcc
     6361 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     6421 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     6481 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     6541 atgctgggga tgcggtgggc tctatggcag gaacccctag tgatggagtt ggccactccc
     6601 tctctgcgcg ctcgct
//
