LOCUS       sequence-325806-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing tTA2 AAV Virus. Alias: AiP1011 - 
            pAAV-AiE2004m-minBG-tTA2-WPRE-HGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  AUTHORS   Graybuck LT, Daigle TL, Sedeno-Cortes AE, Walker M, Kalmbach B, Lenz
            GH, Morin E, Nguyen TN, Garren E, Bendrick JL, Kim TK, Zhou T, 
            Mortrud M, Yao S, Siverts LA, Larsen R, Gore BB, Szelenyi ER, Trader
            C, Balaram P, van Velthoven CTJ, Chiang M, Mich JK, Dee N, Goldy J, 
            Cetin AH, Smith K, Way SW, Esposito L, Yao Z, Gradinaru V, Sunkin 
            SM, Lein E, Levi BP, Ting JT, Zeng H, Tasic B
  TITLE     Enhancer viruses for combinatorial cell-subclass-specific labeling.
  JOURNAL   Neuron. 2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011.
  PUBMED    33789083
REFERENCE   2  (bases 1 to 5418)
  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: "Neuron. 
            2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011."
COMMENT     Sequence Label: AiP1011-pAAV-mscRE4-minBGpromoter-tTA2-WPRE-hGHpA
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(774..1391,1392..1397,1398..1433,1434..1436,1437..1472,
                     1473..1475,1476..1511,1512..1520)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    1545..2133
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1598..1618)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2016..2027)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2165..2641
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2364..2383)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2681..2821
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2681..2810
                     /label=AAV2 ITR
     rep_origin      2896..3351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2983..3002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3193..3214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3368..3387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3487..3509
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3547..3565)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3633..3737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3738..3806,3807..4598)
                     /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(3956..3975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4769..5357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5258..5277
                     /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 gcgtataagc cttgggggca atcaaactat tacattgagt
      181 ccttggattt gctacaaatt acattttaaa tgcaatcatt ttataaaagc ttcaacactc
      241 acacttggaa gcgttaccct gttgaatatc actgactcac taacttgcat tgccatgcta
      301 acttgctttc agagagatct cagaacacat catcttctgc tatttcaata catgcacatt
      361 aatttcctat caacgtgtgc tgatcaggaa ctctgtaatc tggcaccggt gtttattttt
      421 attcctgtct attcctgttg gctcacgaaa agattgtttg agcaagtgtt ttatggtgag
      481 ttgtatcata tgtacattga tttaatctgc ccacattcag ttctacaagc ggagccaaaa
      541 aaatagagac aagcataatt ttcattcaac atgagcccct caatgcaagc caagtacctc
      601 atctggtgct cagctaaagc aacagcaatc tgttccaccc tggagacaca actggccaca
      661 gaaaacttag tgaaaagagg caatgctatg cacaggacaa atgagctcgg gctgggcata
      721 aaagtcaggg cagagccatc tattgcttac atttgcttct gggatccgcc accatgtcta
      781 gactggacaa gagcaaagtc ataaactctg ctctggaatt actcaatgaa gtcggtatcg
      841 aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact
      901 ggcacgtgaa gaacaagcgg gccctgctcg atgccctggc aatcgagatg ctggacaggc
      961 atcataccca cttctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg
     1021 ccaagtcatt ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca
     1081 cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc
     1141 aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg
     1201 gctgcgtatt ggaggatcag gagcatcaag tagcaaaaga ggaaagagag acacctacca
     1261 ccgattctat gcccccactt ctgagacaag caattgagct gttcgaccat cagggagccg
     1321 aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt
     1381 gcgaaagcgg cgggccggcc gacgcccttg acgattttga cttagacatg ctcccagccg
     1441 atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc
     1501 ttgacatgct ccccgggtaa gaattcgata tcaagcttat cgataatcaa cctctggatt
     1561 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     1621 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     1681 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     1741 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     1801 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     1861 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     1921 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     1981 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2041 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2101 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     2161 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     2221 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     2281 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     2341 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     2401 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     2461 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     2521 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     2581 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     2641 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     2701 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2761 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2821 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     2881 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2941 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3001 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3061 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3121 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3181 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     3241 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     3301 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     3361 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     3421 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3481 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3541 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3601 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3661 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5401 ggccttttgc tcacatgt
//
