LOCUS       sequence-385570-        5643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector for astrocyte-selective Cre-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 5643)
  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 5643)
  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-flex-MCS--4x6T
FEATURES             Location/Qualifiers
     source          1..5643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(145..936,937..1005))
                     /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     768..787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1006..1110)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1136..1591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1273..1294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1485..1504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1663..1792
                     /label=AAV2 ITR
     promoter        2103..2380
                     /label=chicken beta-actin promoter
     primer_bind     2659..2681
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2743..2762
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2797..2830
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2893..2926
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3570..3603)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(3674..3707)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3749..4337
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3802..3822)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4220..4231)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4380..4501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4391..4410)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4445..4464
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4663..4792
                     /label=AAV2 ITR
     rep_origin      complement(5029..5617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5109..5128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
       61 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
      241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
      361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
      421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
      481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
      541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
      601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
      661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
      721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
      781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
      841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     1141 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1201 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1261 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1321 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     1381 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1441 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1501 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     1561 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     1621 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc
     1681 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     1741 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta
     1801 atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagag taccattgac
     1861 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     1921 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1981 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     2041 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2101 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     2161 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     2221 ggggcgcgcg ccaggsgggg sggggsgggg sgaggggsgg ggsggggsga ggsggagagg
     2281 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     2341 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     2401 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     2461 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     2521 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     2581 gggccctttg tgcgggggga gcggctcggg gctgtccgcg gggggacggc tgccttcggg
     2641 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     2701 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     2761 tgctgtctca tcattttggc aaagaattgg atccccataa cttcgtataa agtatcctat
     2821 acgaagttat atcaaaatag gaagaccaat gcttcaccat cgacccgaat tgccaagcat
     2881 caccatcgac ccataacttc gtataatgta tgctatacga agttatacta gggatccccc
     2941 cgtcgcggcc gctttacctg gtacccaggt cgtcttgcag ggcttctaac acacccagct
     3001 aaccttttta gatcgaccgt gttatattcg cttattgctt aagaatacgc gtagtaaggc
     3061 acgcggtgaa tgcctcgtac cgtgagtaat aatgcgtcgt accgtgagta ataatgcggt
     3121 acccaggtcg tcttgcaggg cttctaacac acccagctaa cctttttaga tcgaccgtgt
     3181 tatattcgct tattgcttaa gaatacgcgt agtaaggcac gcggtgaatg cccaggtcgt
     3241 cttgcagggc ttctaacaca cccagctaac ctttttagat cgaccgtgtt atattcgctt
     3301 attgcttaag aatacgcgta gtaaggcacg cggtgaatgc ctcgtaccgt gagtaataat
     3361 gcgtcgtacc gtgagtaata atgcgcaggt cgtcttgcag ggcttctaac acacccagct
     3421 aaccttttta gatcgaccgt gttatattcg cttattgctt aagaatacgc gtagtaaggc
     3481 acgcggtgaa tgcccactcg agcacgtggc tagcgagctc accggtttcg aactgcagat
     3541 gcattcttac gcgtgtaccg aattctagta taacttcgta taggatactt tatacgaagt
     3601 tatcattggg attcttccta ttttgatcca agcatcacca tcgaccctct agtccagatc
     3661 tcaccatcga cccataactt cgtatagcat acattatacg aagttatgtc cctcgaagag
     3721 gttcgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3781 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3841 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3901 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3961 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4021 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4081 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4141 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4201 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4261 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4321 ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat
     4381 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4441 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4501 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4561 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
     4621 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     4681 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4741 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta
     4801 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4861 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4921 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4981 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5041 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5101 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5161 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5221 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5281 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5341 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5401 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5461 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5521 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5581 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5641 ggg
//
