LOCUS       sequence-385348-        6829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dre-dependent AAV expression of membrane-targeted Myc spaghetti 
            monster reporter preferentially in astrocytes; astrocyte selectivity
            generated with 4x6T miRNA targeting cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  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 6829)
  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-lck-smMyc-4x6T
FEATURES             Location/Qualifiers
     source          1..6829
                     /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"
     CDS             complement(3559..3588)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3592..3621)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3625..3654)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3868..3897)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3904..3933)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3946..3975)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3982..4011)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(4543..4572)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(4576..4605)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(4609..4638)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      4638..4647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4779..4788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4805..4836
                     /label=rox
                     /bound_moiety="Dre recombinase"
     misc_feature    4961..5549
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5014..5034)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5432..5443)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5592..5713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5603..5622)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5657..5676
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5875..6004
                     /label=AAV2 ITR
     rep_origin      complement(6241..6829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6321..6340)
                     /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 gagaggkgcg 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 cccatatgct aatcgttaca gatcctcttc gctgataagt ttctgctccg ccaggtcctc
     3601 ttcgctgatc agcttctgtt ccgccagatc ctcctcggag ataagtttct gttcagcgcc
     3661 cttgtacagt tcgtccatgc ccaacgtgat gcccgcagcc gtgacgaact caagaagaac
     3721 catgtggtca cgtttttcat tcgggtcctt gctcagcacc gactgggtgg agaggtagtg
     3781 gttatccggc agaaggacgg ggccgtcacc aattggcgta ttctgctggt agtgatccgc
     3841 caactggaca cttccatctc ctcccgcgag atcctcctcc gagatgagct tttgctcggc
     3901 cgccaggtct tcctccgata tcagcttctg ctcgccgcct ccgcccaagt cctcctcgct
     3961 aatgagtttc tgctctgcgg ccagatcctc ttcactgatg agcttctgtt ctgcgccgcc
     4021 ttcgacattg tgacgaattt tgaaattggc tttaatgcca ttcttctgct tatcggcagt
     4081 tatatagaca ttgtgactgt tgaagttata ctccaacttg tggcccagga tattgccatc
     4141 ctccttaaaa tctatgccct tcagctcgat acgattcacg agtgtatcac cctcgaattt
     4201 cacctcagcc cgtgtcttgt aggtgccgtc gtccttgaaa ctgatggtcc tctcttgaac
     4261 gtaaccctcg ggcatagcgc tcttgaaaaa gtcatgctgc ttcatgtgat ctggataccg
     4321 cgaaaaacac tgaactccgc cacccaaggt tgtgaccagg gtgggccagg gtacgggcag
     4381 tttacccgtg gtgcaaatga acttcagggt cagtttaccg ttggttgcgt cgccctcgcc
     4441 ctctccccgt acggagaatt tatgtccatt gacatctccg tccagctcga cgaggatggg
     4501 caccacgccg gtgaacagct cttcaccctt cgacactccg gcaaggtctt cctcactgat
     4561 cagtttttgt tcggcaagat cctcttccga aatcagtttc tgctctgcca agtcttcctc
     4621 cgatatcagt ttttgctcca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     4681 ctgcagaatt cgaagcttcg gcgggggata atggcagttc tcacacacgt caatgttctc
     4741 catccagtca tcttcagggt ttgagctgca gccacagccc atggtggcag atctgagggc
     4801 tagctaactt taaataattg gcattattta aagttattgc cttaacccag aaattatcac
     4861 tgttattctt tagaatggtg caaagataac tttaaataat cgacattatt taaagttaga
     4921 attcgatatc aagcttatcg ataatatcaa gcttatcgat aatcaacctc tggattacaa
     4981 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5041 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5101 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5161 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5221 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5281 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5341 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     5401 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5461 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5521 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgcttcg
     5581 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     5641 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     5701 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     5761 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct
     5821 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     5881 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     5941 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     6001 gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6061 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6241 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6661 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6781 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaa
//
