LOCUS       sequence-385569-        5614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector for astrocyte-selective Dre-dependent AAV expression; 
            astrocyte selectivity generated with 4x6T miRNA targeting cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  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 5614)
  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-CAG-dDIO-MCS--4x6T
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1689..1818
                     /label=AAV2 ITR
     promoter        2129..2406
                     /label=chicken beta-actin promoter
     primer_bind     2685..2707
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2769..2788
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2865..2896
                     /label=rox
                     /bound_moiety="Dre recombinase"
     protein_bind    3590..3621
                     /label=rox
                     /bound_moiety="Dre recombinase"
     misc_feature    3746..4334
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3799..3819)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4217..4228)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4377..4498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4388..4407)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4442..4461
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4660..4789
                     /label=AAV2 ITR
     rep_origin      complement(5026..5614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5106..5125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     1741 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     1801 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
     1861 catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1921 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1981 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2041 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2101 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
     2161 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
     2221 gcagcgatgg gggcgggggg gggggggggg cgcgcgccrg gsggggsggg gsggggsgrg
     2281 gggsggggsg gggsgrggcg grgaggtgcg gcggcagcca atcagagcgg cgcgctccga
     2341 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
     2401 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
     2461 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
     2521 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
     2581 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg
     2641 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
     2701 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
     2761 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc
     2821 ctagtccggc tagaggtacc ggatcctcta gagtcgactc cggataactt taaataatgc
     2881 caattattta aagttagcag aatggtagct ggattgtagc tgctattagc aatatgaaac
     2941 ctcttataac tttaaataat gtcgattatt taaagttagg cgcgccctag tcgcggccgc
     3001 tttacctggt acccaggtcg tcttgcaggg cttctaacac acccagctaa cctttttaga
     3061 tcgaccgtgt tatattcgct tattgcttaa gaatacgcgt agtaaggcac gcggtgaatg
     3121 cctcgtaccg tgagtaataa tgcgtcgtac cgtgagtaat aatgcggtac ccaggtcgtc
     3181 ttgcagggct tctaacacac ccagctaacc tttttagatc gaccgtgtta tattcgctta
     3241 ttgcttaaga atacgcgtag taaggcacgc ggtgaatgcc caggtcgtct tgcagggctt
     3301 ctaacacacc cagctaacct ttttagatcg accgtgttat attcgcttat tgcttaagaa
     3361 tacgcgtagt aaggcacgcg gtgaatgcct cgtaccgtga gtaataatgc gtcgtaccgt
     3421 gagtaataat gcgcaggtcg tcttgcaggg cttctaacac acccagctaa cctttttaga
     3481 tcgaccgtgt tatattcgct tattgcttaa gaatacgcgt agtaaggcac gcggtgaatg
     3541 cccaagatct cacgtgaccg gtgctagcga gctcttaatt aagcatgcct aactttaaat
     3601 aattggcatt atttaaagtt attgccttaa cccagaaatt atcactgtta ttctttagaa
     3661 tggtgcaaag ataactttaa ataatcgaca ttatttaaag ttagaattcg atatcaagct
     3721 tatcgataat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     3781 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3841 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3901 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3961 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4021 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4081 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4141 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4201 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4261 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4321 ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag
     4381 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4441 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4501 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     4561 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     4621 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     4681 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4741 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg
     4801 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4861 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4921 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4981 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5221 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5461 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5581 gcagcagatt acgcgcagaa aaaaaggatc tcaa
//
