LOCUS       sequence-385067-        6855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Astrocytic expression of light-inducible iCreV 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 6855)
  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 6855)
  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-iCreV-4x6T
FEATURES             Location/Qualifiers
     source          1..6855
                     /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      1527..1536
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2136..2156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2172..2228
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2238..2258
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4268..4744
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4467..4486)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4790..4930
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4790..4919
                     /label=AAV2 ITR
     rep_origin      5005..5460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5092..5111)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5302..5323
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5477..5496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5596..5618
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5656..5674)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5742..5846
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5847..5915,5916..6707)
                     /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 aattcgatct taagccgcca ccatggccac ctctgatgaa gtcaggaaga
     1561 acctgatgga catgttcagg gacaggcagg ccttctctga acacacctgg aagatgctcc
     1621 tgtctgtgtg cagatcctgg gctgcctggt gcaagctgaa cggtacccat acactgtatg
     1681 cccctggagg atatgatatt atgggatatc tgattcagat tatgaaaaga cctaatcctc
     1741 aggtggaact gggacctgtg gatacaagcg tggccctgat tctgtgtgat ctgaaacaga
     1801 aagatacacc tattgtgtat gccagcgaag cctttctgta tatgacagga tatagcaatg
     1861 ccgaagtgct gggaagaaat tgtagatttc tgcagagccc tgatggaatg gtgaaaccta
     1921 aaagcacaag aaaatatgtg gatagcaata caattaatac aatgagaaaa gccattgata
     1981 gaaatgccga agtgcaggtg gaagtggtga attttaaaaa aaatggacag agatttgtga
     2041 attttctgac aatgattcct gtgagagatg aaacaggaga atatagatat agcatgggat
     2101 ttcagtgtga aacagaaggc ggaagcggtg gcgtgcccaa gaagaagagg aaagtcggat
     2161 ccggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc
     2221 ccggccccct cgaggtgccc aagaagaaga ggaaagtcgg cggacatact ctgtacgctc
     2281 ccggagggta tgacataatg ggttacctaa tccagattat gaagcgtcct aatccccagg
     2341 ttgagttggg accagtggac accagcgtag ccttaattct ttgcgacctt aaacagaaag
     2401 atacaccaat agtatacgcg agtgaggctt ttttgtatat gacgggttat tccaacgcgg
     2461 aagtgttagg acgaaattgt cggtttctac aatcaccgga tgggatggtc aaaccgaaaa
     2521 gtactagaaa gtatgtggat tcgaatacaa ttaacaccat gagaaaggca atcgatagga
     2581 acgcagaagt acaagtcgag gttgtcaatt tcaaaaagaa tgggcaacgc ttcgtaaact
     2641 ttctcactat gatacctgtt agggacgaaa cgggcgagta tcgatactct atgggcttcc
     2701 aatgtgaaac agaaggtacc aacaggaaat ggttccctgc tgaacctgag gatgtgaggg
     2761 actacctcct gtacctgcaa gccagaggcc tggctgtgaa gaccatccaa cagcacctgg
     2821 gccagctcaa catgctgcac aggagatccg gcctgcctcg cccttctgac tccaatgctg
     2881 tgtccctggt gatgaggaga atcagaaagg agaatgtgga tgctggggag agagccaagc
     2941 aggccctggc ctttgaacgc actgactttg accaagtcag atccctgatg gagaactctg
     3001 acagatgcca ggacatcagg aacctggcct tcctgggcat tgcctacaac accctgctgc
     3061 gcattgccga aattgccaga atcagagtga aggacatctc ccgcaccgat ggtgggagaa
     3121 tgctgatcca cattggcagg accaagaccc tggtgtccac agctggtgtg gagaaggccc
     3181 tgtccctggg ggttaccaag ctggtggaga gatggatctc tgtgtctggt gtggctgatg
     3241 accccaacaa ctacctgttc tgccgggtca gaaagaatgg tgtggctgcc ccttctgcca
     3301 cctcccaact gtccacccgg gccctggaag ggatctttga ggccacccac cgcctgatct
     3361 atggtgccaa ggatgactct gggcagagat acctggcctg gtctggccac tctgccagag
     3421 tgggtgctgc cagggacatg gccagggctg gtgtgtccat ccctgaaatc atgcaggctg
     3481 gtggctggac caatgtgaac attgtgatga actacatcag aaacctggac tctgagactg
     3541 gggccatggt gaggctgctc gaagatgggg actaacatat gggcattcac cgcgtgcctt
     3601 actacgcgta ttcttaagca ataagcgaat ataacacggt cgatctaaaa aggttagctg
     3661 ggtgtgttag aagccctgca agacgacctg cgcattatta ctcacggtac gacgcattat
     3721 tactcacggt acgaggcatt caccgcgtgc cttactacgc gtattcttaa gcaataagcg
     3781 aatataacac ggtcgatcta aaaaggttag ctgggtgtgt tagaagccct gcaagacgac
     3841 ctgggcattc accgcgtgcc ttactacgcg tattcttaag caataagcga atataacacg
     3901 gtcgatctaa aaaggttagc tgggtgtgtt agaagccctg caagacgacc tgggtaccgc
     3961 attattactc acggtacgac gcattattac tcacggtacg aggcattcac cgcgtgcctt
     4021 actacgcgta ttcttaagca ataagcgaat ataacacggt cgatctaaaa aggttagctg
     4081 ggtgtgttag aagccctgca agacgacctg ggtaccaggt aaagcggccg cgactctaga
     4141 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4201 tccccctgaa cctgaaacat aaaatgaatg caattctgcc tcgagcagcg ctgctcgaga
     4261 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     4321 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     4381 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     4441 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     4501 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     4561 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     4621 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     4681 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     4741 ccttctgatt ttgtaggtaa ccacgtggtg gtgcggaccg agcggccgca ggaaccccta
     4801 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4861 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4921 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4981 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5041 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     5101 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5161 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5221 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5281 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     5341 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     5401 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5461 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5521 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5581 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5641 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5701 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5761 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5821 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5881 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5941 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6001 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6061 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6121 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6181 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6241 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6301 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6361 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6421 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6481 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6541 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6601 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6661 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6721 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6781 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6841 gagcgtcaga ccccg
//
