LOCUS       sequence-385152-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Astrocytic expression of Dre recombinase in AAV; astrocyte 
            specificity enhanced with 4x6T miRNA targeting cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  AUTHORS   Gleichman AJ, Kawaguchi R, Sofroniew MV, Carmichael ST
  TITLE     A toolbox of astrocyte-specific, serotype-independent 
            adeno-associated viral vectors using microRNA targeting sequences.
  JOURNAL   Nat Commun. 2023 Nov 16;14(1):7426. doi: 10.1038/s41467-023-42746-w.
  PUBMED    37973910
REFERENCE   2  (bases 1 to 5863)
  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 
            Commun."; date: "2023-11-16"; volume: "14(1):7426. doi"; pages: " 
            10.1038/s41467-023-42746-w"
COMMENT     Sequence Label: AAV-GfaABC1D-DreO-4x6T
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   673..802
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1532..1552
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1553..2575
                     /product="site-specific recombinase"
                     /label=Dre recombinase
                     /note="Dre is a site-specific recombinase from 
                     bacteriophage D6. Recombination occurs at rox sequences."
     polyA_signal    3276..3752
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3475..3494)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3798..3938
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3798..3927
                     /label=AAV2 ITR
     rep_origin      4013..4468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4100..4119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4310..4331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4485..4504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4604..4626
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4664..4682)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4750..4854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4855..4923,4924..5715)
                     /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"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
      721 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      781 aactccatca ctaggggttc ctgcggccgc acgcgtacat atcctggtgt ggagtagggg
      841 acgctgctct gacagaggct cgggggcctg agctggctct gtgagctggg gaggaggcag
      901 acagccaggc cttgtctgca agcagacctg gcagcattgg gctggccgcc ccccagggcc
      961 tcctcttcat gcccagtgaa tgactcacct tggcacagac acaatgttcg gggtgggcac
     1021 agtgcctgct tcccgccgca ccccagcccc cctcaaatgc cttccgagaa gcccattgag
     1081 cagggggctt gcattgcacc ccagcctgac agcctggcat cttgggataa aagcagcaca
     1141 gccccctagg ggctgccctt gctgtgtggc gccaccggcg gtggagaaca aggctctatt
     1201 cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc atggacagtg ggtggcaggg
     1261 ggggagagga gggctgtctg cttcccagaa gtccaaggac acaaatgggt gaggggagag
     1321 ctctccccat agctgggctg cggcccaacc ccaccccctc aggctatgcc agggggtgtt
     1381 gccaggggca cccgggcatc gccagtctag cccactcctt cataaagccc tcgcatccca
     1441 ggagcgagca gagccagagc aggttggaga ggagacgcat cacctccgct gctcgcttaa
     1501 ttaatacctg aattcatcga ttgccaccat gcctaagaag aagaggaagg tttctgagct
     1561 gattattagt ggttcatctg gtggattcct gcgaaacatc ggcaaagagt atcaggaggc
     1621 cgctgaaaac ttcatgaggt ttatgaatga ccagggggcg tacgctccta acactttgag
     1681 ggatttgagg ttggtctttc atagctgggc cagatggtgc catgctcggc agcttgcatg
     1741 gtttccaatt agtcctgaaa tggcacgcga atactttctt cagttgcacg atgcagacct
     1801 ggcctccact accatcgaca agcactatgc tatgcttaat atgcttctgt cccactgcgg
     1861 actgccaccc ttgtccgacg acaagtcagt gagtcttgcc atgagaagaa ttagaagaga
     1921 agccgcaacc gaaaagggtg agaggacagg acaggcaatc cccctgcgct gggacgacct
     1981 gaagctgctg gatgtgctgc tcagcaggag cgagcggctg gtcgacctgc gcaacagggc
     2041 tttcctgttc gtagcctata acaccctcat gagaatgtct gaaatatcac gcatcagggt
     2101 tggggacttg gatcagacag gagacacagt gaccctgcac atcagtcaca ctaagacaat
     2161 caccacagct gcgggccttg acaaagtgct ctcccggcga accacagcag tgctcaatga
     2221 ctggctggac gtcagtgggc ttagagaaca tccagacgct gtgctcttcc cacctataca
     2281 ccggtcaaac aaagcccgca ttactaccac gcccctgacc gcccctgcca tggagaagat
     2341 tttcagtgat gcctgggtgc tgctgaacaa acgggacgcc acccccaata aagggaggta
     2401 taggacctgg accggccatt ccgccagggt gggtgccgca atagacatgg ccgagaaaca
     2461 ggtgtctatg gtcgagatta tgcaggaagg gacatggaag aagcctgaaa cactgatgcg
     2521 gtatctcaga aggggcggag tgtccgtggg agccaattct cgactgatgg atagctaagg
     2581 atccatatgg gcattcaccg cgtgccttac tacgcgtatt cttaagcaat aagcgaatat
     2641 aacacggtcg atctaaaaag gttagctggg tgtgttagaa gccctgcaag acgacctgcg
     2701 cattattact cacggtacga cgcattatta ctcacggtac gaggcattca ccgcgtgcct
     2761 tactacgcgt attcttaagc aataagcgaa tataacacgg tcgatctaaa aaggttagct
     2821 gggtgtgtta gaagccctgc aagacgacct gggcattcac cgcgtgcctt actacgcgta
     2881 ttcttaagca ataagcgaat ataacacggt cgatctaaaa aggttagctg ggtgtgttag
     2941 aagccctgca agacgacctg ggtaccgcat tattactcac ggtacgacgc attattactc
     3001 acggtacgag gcattcaccg cgtgccttac tacgcgtatt cttaagcaat aagcgaatat
     3061 aacacggtcg atctaaaaag gttagctggg tgtgttagaa gccctgcaag acgacctggg
     3121 taccaggtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     3181 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3241 attctgcctc gagcagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     3301 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     3361 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3421 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3481 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3541 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     3601 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     3661 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     3721 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtggtggt
     3781 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3841 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     3901 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     3961 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4021 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4081 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4141 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4201 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4261 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4321 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4381 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4441 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4501 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4561 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4621 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4681 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4741 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4801 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4861 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4921 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4981 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5041 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5101 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5161 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5221 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5281 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5341 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5401 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5461 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5521 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5581 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5641 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5701 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5761 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5821 atcccttaac gtgagttttc gttccactga gcgtcagacc ccg
//
