LOCUS       sequence-329446-        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"
     promoter        126..573
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    613..631
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    649..667
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    770..803
                     /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    854..901
                     /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(932..1042)
                     /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(1680..1713)
                     /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(1778..1825)
                     /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    1846..2434
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1899..1919)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2317..2328)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2466..2942
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2665..2684)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2982..3122
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2982..3111
                     /label=AAV2 ITR
     rep_origin      3197..3652
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3284..3303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3494..3515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3669..3688)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3788..3810
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3848..3866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3934..4038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4039..4107,4108..4899)
                     /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(4257..4276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5070..5658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5559..5578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5720..5768,1..81)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
       61 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      121 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      181 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      241 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      301 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      361 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      421 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      481 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      541 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa cagatctcta aaacaggtaa
      601 gtcccattaa tctccctatc agtgatagag aaggtctgaa gagtttactc cctatcagtg
      661 atagagatta attcctctac taaccttgtt catcttttct ttttttttct acaggtcctg
      721 ggtgattaac agcttaaggg cccggatccg atatcgaagt tcctattccg aagttcctat
      781 tcttcaaata gtataggaac ttcgcagaat ggtagctgga ttgtagctgc tattagcaat
      841 atgaaacctc ttagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt
      901 cgagctcggc gcgccggaat tcttacccgg ggagcatgtc aaggtcaaaa tcgtcaagag
      961 cgtcagcagg cagcatatca aggtcaaagt cgtcaagggc atcggctggg agcatgtcta
     1021 agtcaaaatc gtcaagggcg tcggccggcc cgccgctttc gcactttagc tgtttctcca
     1081 ggccacatat gattagttcc aggccgaaaa ggaaggcagg ttcggctccc tgccggtcga
     1141 acagctcaat tgcttgtctc agaagtgggg gcatagaatc ggtggtaggt gtctctcttt
     1201 cctcttttgc tacttgatgc tcctgttcct ccaatacgca gcccagtgta aagtggccca
     1261 cggcggacag agcgtacagt gcgttctcca gggagaagcc ttgctgacac aggaacgcga
     1321 gctgattttc cagggtttcg tactgtttct ctgttgggcg ggtgccgaga tgcactttag
     1381 ccccgtcgcg atgtgagagg agagcacagc ggaatgactt ggcgttgttc cgcagaaagt
     1441 cttgccatga ctcgccttcc agggggcaga agtgggtatg atgcctgtcc agcatctcga
     1501 tggccagggc atcgagcagg gcccgcttgt tcttcacgtg ccagtacagg gtaggctgct
     1561 caactcccag cttttgagcg agtttccttg tcgtcaggcc ttcgataccg acttcattga
     1621 gtaattccag agcgccgttt atgactttag atttatccag ccgactcatg tcgacccggg
     1681 aagttcctat actatttgaa gaataggaac ttcggaatag gaacttcttg ccttaaccca
     1741 gaaattatca ctgttattct ttagaatggt gcaaagagaa gttcctatac tttctagaga
     1801 ataggaactt cggaatagga acttcgaagt actaagctta tcgataatca acctctggat
     1861 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1921 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     1981 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2041 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2101 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2161 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2221 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     2281 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2341 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2401 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     2461 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     2521 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     2581 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     2641 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     2701 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     2761 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     2821 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     2881 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     2941 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3001 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3061 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3121 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3181 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3241 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     3301 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3361 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3421 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3481 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3541 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     3601 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3661 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3721 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3781 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3841 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3901 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3961 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4021 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4081 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4141 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4201 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4261 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4321 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4381 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4441 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4501 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4561 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4621 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4681 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4741 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4801 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4861 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4921 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4981 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5701 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
     5761 cgggcgtc
//
