LOCUS       sequence-325808-        5443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing tTA2 AAV Virus. Alias: AiP1049 - 
            pAAV-AiE2013m-minBG-tTA2-WPRE-HGHpA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5443)
  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 5443)
  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: AiP1049-pAAV-mscRE13-minBGpromoter-tTA2-WPRE-h...
FEATURES             Location/Qualifiers
     source          1..5443
                     /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(799..1416,1417..1422,1423..1458,1459..1461,1462..1497,
                     1498..1500,1501..1536,1537..1545)
                     /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    1570..2158
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1623..1643)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2041..2052)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2190..2666
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2389..2408)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2706..2846
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2706..2835
                     /label=AAV2 ITR
     rep_origin      2921..3376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3008..3027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3218..3239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3393..3412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3512..3534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3572..3590)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3658..3762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3763..3831,3832..4623)
                     /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(3981..4000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4794..5382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5283..5302
                     /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 gcgtgtccca taggcagttt gtggctgagt gctggtccag
      181 gggtgaggag gtgggctatg ttactgaggg tctctgggtg ttaggaaaac agggcccagg
      241 agtctggctg ctcgtatgct ggcccaggct cttgtttttc ttgagctgac ttgctggaga
      301 agtgagctaa gtcagaaaca aaatgccaca ttgcacgccc actgaagtct gggctcaagg
      361 gaaagaagag agattgccag agcgttagct gttcccaatc cactcctgga ccttaagctg
      421 tcttgaacag agttgccaat cagcttggta gggactggcc tttgaggagg ggagggggtg
      481 taggcagggg agggggagag aagggagcag tctgcgctcc atcttaatta cctcatcaga
      541 aacagctccc ttcccgcaaa gctctggtgt cttctacaag agggtgagtc tttggcttta
      601 catgtgaact tgtgccattt gcctgcgtat ataaacatga agggtcgtct gggttcagag
      661 ctgaaatctt tcacttgtga cttagctggg aaattcttgg caagatcaga atgcagtggt
      721 aaggtctgag ctcgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg
      781 cttctgggat ccgccaccat gtctagactg gacaagagca aagtcataaa ctctgctctg
      841 gaattactca atgaagtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
      901 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
      961 ctggcaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     1021 tggcaagact ttctgcggaa caacgccaag tcattccgct gtgctctcct ctcacatcgc
     1081 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     1141 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     1201 tccgccgtgg gccactttac actgggctgc gtattggagg atcaggagca tcaagtagca
     1261 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     1321 gagctgttcg accatcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     1381 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cggccgacgc ccttgacgat
     1441 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     1501 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaagaatt cgatatcaag
     1561 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1621 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1681 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     1741 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     1801 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     1861 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1921 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     1981 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2041 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2101 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     2161 cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     2221 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     2281 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     2341 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     2401 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     2461 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     2521 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     2581 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     2641 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     2701 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     2761 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     2821 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     2881 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     2941 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     3001 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3061 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3121 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3181 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3241 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     3301 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3361 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3421 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3481 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3541 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3601 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3661 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3721 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3781 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3841 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3901 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3961 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4021 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4081 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4141 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4201 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4261 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4321 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4381 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4441 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4501 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4561 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4621 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4681 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4741 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4861 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4921 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4981 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5041 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5101 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5161 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5221 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5281 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5341 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5401 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
