LOCUS       sequence-323219-        5768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Flp-dependent tTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5768)
  AUTHORS   Sabatini PV, Wang J, Rupp AC, Affinati AH, Flak JN, Li C, Olson DP, 
            Myers MG Jr
  TITLE     tTARGIT AAVs mediate the sensitive and flexible manipulation of 
            intersectional neuronal populations in mice.
  JOURNAL   eLife. 2021 Mar 11;10. pii: 66835. doi: 10.7554/eLife.66835.
  PUBMED    33704065
REFERENCE   2  (bases 1 to 5768)
  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: "eLife."; 
            date: "2021-03-11"; volume: "10. pii: 66835. doi"; pages: " 
            10.7554/eLife.66835"
COMMENT     Sequence Label: pAAV-hSYN1-fDIO-tTA
FEATURES             Location/Qualifiers
     source          1..5768
                     /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    819..852
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    903..950
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(981..1091)
                     /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    complement(1729..1762)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    complement(1827..1874)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    1895..2483
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1948..1968)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2366..2377)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2515..2991
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2714..2733)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3031..3171
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3031..3160
                     /label=AAV2 ITR
     rep_origin      3246..3701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3333..3352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3543..3564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3718..3737)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3837..3859
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3897..3915)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3983..4087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4088..4156,4157..4948)
                     /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(4306..4325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5119..5707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5608..5627
                     /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 ccggatccga tatcgaagtt cctattccga agttcctatt cttcaaatag
      841 tataggaact tcgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct
      901 tagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc gagctcggcg
      961 cgccggaatt cttacccggg gagcatgtca aggtcaaaat cgtcaagagc gtcagcaggc
     1021 agcatatcaa ggtcaaagtc gtcaagggca tcggctggga gcatgtctaa gtcaaaatcg
     1081 tcaagggcgt cggccggccc gccgctttcg cactttagct gtttctccag gccacatatg
     1141 attagttcca ggccgaaaag gaaggcaggt tcggctccct gccggtcgaa cagctcaatt
     1201 gcttgtctca gaagtggggg catagaatcg gtggtaggtg tctctctttc ctcttttgct
     1261 acttgatgct cctgttcctc caatacgcag cccagtgtaa agtggcccac ggcggacaga
     1321 gcgtacagtg cgttctccag ggagaagcct tgctgacaca ggaacgcgag ctgattttcc
     1381 agggtttcgt actgtttctc tgttgggcgg gtgccgagat gcactttagc cccgtcgcga
     1441 tgtgagagga gagcacagcg gaatgacttg gcgttgttcc gcagaaagtc ttgccatgac
     1501 tcgccttcca gggggcagaa gtgggtatga tgcctgtcca gcatctcgat ggccagggca
     1561 tcgagcaggg cccgcttgtt cttcacgtgc cagtacaggg taggctgctc aactcccagc
     1621 ttttgagcga gtttccttgt cgtcaggcct tcgataccga cttcattgag taattccaga
     1681 gcgccgttta tgactttaga tttatccagc cgactcatgt cgacccggga agttcctata
     1741 ctatttgaag aataggaact tcggaatagg aacttcttgc cttaacccag aaattatcac
     1801 tgttattctt tagaatggtg caaagagaag ttcctatact ttctagagaa taggaacttc
     1861 ggaataggaa cttcgaagta ctaagcttat cgataatcaa cctctggatt acaaaatttg
     1921 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1981 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2041 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2101 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2161 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2221 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2281 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     2341 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2401 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2461 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     2521 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     2581 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     2641 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     2701 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     2761 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     2821 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     2881 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     2941 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     3001 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     3061 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3121 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     3181 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     3241 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3301 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     3361 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3421 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     3481 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3541 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     3601 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     3661 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     3721 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3781 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3841 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3901 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3961 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4021 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4081 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4141 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4201 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4261 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4321 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4381 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4441 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4501 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4561 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4621 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4681 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4741 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4801 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4861 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4921 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4981 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5041 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5101 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5161 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5221 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5281 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5341 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5401 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5461 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5521 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5581 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5641 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5701 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5761 tcacatgt
//
