LOCUS       sequence-351983-        5208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre recombinase dependent tetracycline-controlled 
            transactivator (tTA) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5208)
  AUTHORS   Jin L, Sullivan HA, Zhu M, Lavin TK, Matsuyama M, Fu X, Lea NE, Xu 
            R, Hou Y, Rutigliani L, Pruner M, Babcock KR, Ip JPK, Hu M, Daigle 
            TL, Zeng H, Sur M, Feng G, Wickersham IR
  TITLE     Long-term labeling and imaging of synaptically connected neuronal 
            networks in vivo using double-deletion-mutant rabies viruses.
  JOURNAL   Nat Neurosci. 2024 Jan 11. doi: 10.1038/s41593-023-01545-8.
  PUBMED    38212587
REFERENCE   2  (bases 1 to 5208)
  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: "Nat 
            Neurosci. 2024 Jan 11. doi: 10.1038/s41593-023-01545-8."
COMMENT     Sequence Label: pAAV-syn-FLEX-tTA
FEATURES             Location/Qualifiers
     source          1..5208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   188..328
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   188..317
                     /label=AAV2 ITR
     rep_origin      403..858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(490..509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     700..721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(875..894)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     994..1016
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1054..1072)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1140..1244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1245..1313,1314..2105)
                     /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(1463..1482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2276..2864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2765..2784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2926..3055
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3089..3536
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(3545..3578)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3629..3662)
                     /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)."
     CDS             complement(3816..4436)
                     /codon_start=1
                     /gene="synthetic gene"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     protein_bind    4451..4484
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4535..4568
                     /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)."
     misc_feature    4574..5162
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4627..4647)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5045..5056)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(5170..5208,1..186)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
       61 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
      121 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
      181 ctatggcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
      241 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
      301 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
      361 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
      421 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
      481 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      541 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      601 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      661 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      721 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
      781 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      841 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      901 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      961 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     1021 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     1081 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1141 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1201 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1261 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1321 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1381 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1441 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1501 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1561 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1621 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1681 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1741 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     1801 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     1861 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     1921 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     1981 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2041 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2101 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2161 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2221 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2281 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2341 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2401 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2461 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2521 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2581 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2641 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2701 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2821 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2881 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc
     2941 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca
     3001 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgtcga
     3061 cgctagactg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
     3121 ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca
     3181 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
     3241 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
     3301 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
     3361 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
     3421 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
     3481 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
     3541 attcataact tcgtatagca tacattatac gaagttattc tttgcaccat tctaaagaat
     3601 aacagtgata atttctgggt taaggcaaat aacttcgtat aggatacttt atacgaagtt
     3661 atgctagctc acagcatatc caggtcaaaa tcatcaagtg catcagcagg gagcatgtcc
     3721 aaatcaaagt catccagggc atcagcaggc agcatgtcca ggtcaaaatc atccaaggca
     3781 tcagcaggca cgtgctttct ctttttgggc acgtgagagc cagactcaca tttcagctgc
     3841 ttttccaggc cacagatgat cagttccagg ccaaaaagga atgcaggctc agccccttgg
     3901 tggtcaaaca actcaatagc ttgccgcagc agtggtggca tggagtcagt ggtgggggtc
     3961 tccctctcct ccttggcaac ttggtgctcc tggtcctcca gcacacagcc caaggtgaag
     4021 tggcccacag cagagagggc atacaaggca ttttccaggg agaagccttg ttggcacagg
     4081 aatgccagct ggttctccag ggtctcatac tgcttctctg taggcctggt gcccaagtga
     4141 actttggcac catcccggtg ggagagcaag gcacatctga agctcttggc attgttcctc
     4201 aggaagtcct gccaggactc cccttccaga gggcagaagt gggtgtggtg cctgtccaac
     4261 atctcaatgg ccagggcatc aagcagagcc ctcttgttct tcacatgcca gtacagggta
     4321 ggctgctcca cacccagctt ctgggccagc ttcctggtgg tcaagccctc aatgccaact
     4381 tcattcagga gctccaaggc agagttgatg actttgctct tgtccagtct ggacatggtg
     4441 gcggcgcgcc ataacttcgt ataatgtatg ctatacgaag ttattaagag gtttcatatt
     4501 gctaatagca gctacaatcc agctaccatt ctgcataact tcgtataaag tatcctatac
     4561 gaagttattc cggaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4621 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4681 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4741 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4801 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4861 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4921 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4981 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5041 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5101 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5161 gcctggatcc tgtgccttct agttgccagc catctgttgt ttgcccct
//
