LOCUS       sequence-385068-        6710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Astrocytic expression of membrane-targeted Myc spaghetti monster 
            reporter 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 6710)
  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 6710)
  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-lck-smMyc-4x6T-WPRE
FEATURES             Location/Qualifiers
     source          1..6710
                     /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"
     regulatory      1522..1531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1663..1672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1672..1701
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1705..1734
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1738..1767
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2299..2328
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2335..2364
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2377..2406
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2413..2442
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2656..2685
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2689..2718
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2722..2751
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3474..4062
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3527..3547)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3945..3956)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4120..4596
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4319..4338)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4645..4785
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4645..4774
                     /label=AAV2 ITR
     rep_origin      4860..5315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4947..4966)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5157..5178
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5332..5351)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5451..5473
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5511..5529)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5597..5701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5702..5770,5771..6562)
                     /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 ttaactaccg gactcagatc tgccaccatg ggctgtggct gcagctcaaa ccctgaagat
     1561 gactggatgg agaacattga cgtgtgtgag aactgccatt atcccccgcc gaagcttcga
     1621 attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggagcaaaaa
     1681 ctgatatcgg aggaagactt ggcagagcag aaactgattt cggaagagga tcttgccgaa
     1741 caaaaactga tcagtgagga agaccttgcc ggagtgtcga agggtgaaga gctgttcacc
     1801 ggcgtggtgc ccatcctcgt cgagctggac ggagatgtca atggacataa attctccgta
     1861 cggggagagg gcgagggcga cgcaaccaac ggtaaactga ccctgaagtt catttgcacc
     1921 acgggtaaac tgcccgtacc ctggcccacc ctggtcacaa ccttgggtgg cggagttcag
     1981 tgtttttcgc ggtatccaga tcacatgaag cagcatgact ttttcaagag cgctatgccc
     2041 gagggttacg ttcaagagag gaccatcagt ttcaaggacg acggcaccta caagacacgg
     2101 gctgaggtga aattcgaggg tgatacactc gtgaatcgta tcgagctgaa gggcatagat
     2161 tttaaggagg atggcaatat cctgggccac aagttggagt ataacttcaa cagtcacaat
     2221 gtctatataa ctgccgataa gcagaagaat ggcattaaag ccaatttcaa aattcgtcac
     2281 aatgtcgaag gcggcgcaga acagaagctc atcagtgaag aggatctggc cgcagagcag
     2341 aaactcatta gcgaggagga cttgggcgga ggcggcgagc agaagctgat atcggaggaa
     2401 gacctggcgg ccgagcaaaa gctcatctcg gaggaggatc tcgcgggagg agatggaagt
     2461 gtccagttgg cggatcacta ccagcagaat acgccaattg gtgacggccc cgtccttctg
     2521 ccggataacc actacctctc cacccagtcg gtgctgagca aggacccgaa tgaaaaacgt
     2581 gaccacatgg ttcttcttga gttcgtcacg gctgcgggca tcacgttggg catggacgaa
     2641 ctgtacaagg gcgctgaaca gaaacttatc tccgaggagg atctggcgga acagaagctg
     2701 atcagcgaag aggacctggc ggagcagaaa cttatcagcg aagaggatct gtaaaatagg
     2761 taaagcggcc gcgactctag atcatatggg cattcaccgc gtgccttact acgcgtattc
     2821 ttaagcaata agcgaatata acacggtcga tctaaaaagg ttagctgggt gtgttagaag
     2881 ccctgcaaga cgacctgcgc attattactc acggtacgac gcattattac tcacggtacg
     2941 aggcattcac cgcgtgcctt actacgcgta ttcttaagca ataagcgaat ataacacggt
     3001 cgatctaaaa aggttagctg ggtgtgttag aagccctgca agacgacctg ggcattcacc
     3061 gcgtgcctta ctacgcgtat tcttaagcaa taagcgaata taacacggtc gatctaaaaa
     3121 ggttagctgg gtgtgttaga agccctgcaa gacgacctgg gtaccgcatt attactcacg
     3181 gtacgacgca ttattactca cggtacgagg cattcaccgc gtgccttact acgcgtattc
     3241 ttaagcaata agcgaatata acacggtcga tctaaaaagg ttagctgggt gtgttagaag
     3301 ccctgcaaga cgacctgggt accaggtaaa gcggccgcga ctctagatca taatcagcca
     3361 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3421 gaaacataaa atgaatgcaa ttctgcctcg agcagcgata tcataccgtc gacaatcaac
     3481 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3541 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3601 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3661 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3721 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3781 cggtggaact catcgccgcc tgtcttgccc gctgctggac aggggctcgg ctgttgggca
     3841 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3901 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3961 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4021 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgtctag agtcgacctg
     4081 cagaagcttg cctcgagcag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     4141 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     4201 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     4261 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     4321 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     4381 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     4441 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     4501 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     4561 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgg
     4621 tggtggtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4681 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4741 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4801 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4861 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4921 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4981 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5041 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     5101 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5161 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     5221 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5281 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5341 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5401 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5461 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5521 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5581 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5641 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5701 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5761 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5821 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5881 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5941 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6001 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6061 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6121 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6181 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6241 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6301 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6361 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6421 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6481 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6541 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6601 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6661 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
//
