LOCUS       sequence-418040-        5675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing tetracycline-controlled transactivator (tTA) 
            under Dlx promoter (for expressing tTA in GABAergic neurons).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  AUTHORS   Thompson A, Arano R, Saleem U, Preciado R, Munoz L, Nelson I, Ramos 
            K, Kim Y, Li Y, Xu W
  TITLE     Brain-wide circuit-specific targeting of astrocytes.
  JOURNAL   Cell Rep Methods. 2023 Dec 18;3(12):100653. doi: 
            10.1016/j.crmeth.2023.100653. Epub 2023 Dec 4.
  PUBMED    38052209
REFERENCE   2  (bases 1 to 5675)
  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: "Cell Rep 
            Methods."; date: "2023-12-18"; volume: "3(12):100653. doi"; pages: "
            10.1016/j.crmeth.2023.100653. Epub 2023 Dec 4"
COMMENT     Sequence Label: AAV-Dlx-tTA
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   661..790
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          1509..1641
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1624..1643
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             join(1691..2308,2309..2314,2315..2350,2351..2353,2354..2389,
                     2390..2392,2393..2428,2429..2437)
                     /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    2462..3050
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2515..2535)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2933..2944)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3082..3558
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3281..3300)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3598..3738
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3598..3727
                     /label=AAV2 ITR
     rep_origin      3813..4268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3900..3919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4110..4131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4285..4304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4404..4426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4464..4482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4550..4654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4655..4723,4724..5515)
                     /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(4873..4892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
      721 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      781 aggggttcct gcggccgcac gcgtttattc agaatgttat gcactcacag tggtttggca
      841 tgcatctggt gaattttttt taacgaaaaa ttagtgttgg tttcgatgta tggtagcatt
      901 ctccctaacg taatttgaat aattcagcaa agccccacta ccagctgtac ttctgcagcc
      961 tcttccattc ttttcagcat tataattttg gttaattttc aattttaggt cctacgtctc
     1021 tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct
     1081 gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc
     1141 tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg
     1201 catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt
     1261 ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc
     1321 tacgaaccca aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg
     1381 cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg
     1441 agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca
     1501 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
     1561 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     1621 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taagaaagac
     1681 cggtgccacc atgtctagac tggacaagag caaagtcata aactctgctc tggaattact
     1741 caatgaagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga
     1801 gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctggcaat
     1861 cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt catggcaaga
     1921 ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc gcgacggggc
     1981 taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct
     2041 cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt
     2101 gggccacttt acactgggct gcgtattgga ggatcaggag catcaagtag caaaagagga
     2161 aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa ttgagctgtt
     2221 cgaccatcag ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct
     2281 ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg attttgactt
     2341 agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc
     2401 tcttgacgat tttgaccttg acatgctccc cgggtaagaa ttcgatatca agcttatcga
     2461 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2521 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2581 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2641 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2701 tggttgggga attgccacca cctgtcagct cctttccgga actttcgctt tccccctccc
     2761 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2821 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2881 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2941 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3001 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3061 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     3121 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     3181 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     3241 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     3301 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     3361 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     3421 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3481 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3541 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     3601 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3661 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3721 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3781 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3841 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3901 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3961 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4021 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4081 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4141 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     4201 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4261 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4321 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4381 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4441 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4501 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4561 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4621 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4681 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4741 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4801 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4861 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4921 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4981 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5041 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5101 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5161 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5221 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5281 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5341 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5401 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5461 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5521 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5581 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5641 gttccactga gcgtcagacc ccgtagaaaa gatca
//
