LOCUS       sequence-171076-        5606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of mCherry under the GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5606)
  AUTHORS   Perea G, Yang A, Boyden ES, Sur M
  TITLE     Optogenetic astrocyte activation modulates response selectivity of 
            visual cortex neurons in vivo.
  JOURNAL   Nat Commun. 2014;5:3262. doi: 10.1038/ncomms4262.
  PUBMED    24500276
REFERENCE   2  (bases 1 to 5606)
  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: "2014"; volume: "5:3262. doi"; pages: " 
            10.1038/ncomms4262"
COMMENT     Sequence Label: pAAV-GFAP104-mCherry
FEATURES             Location/Qualifiers
     source          1..5606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..23
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(61..79)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        147..251
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(252..320,321..1112)
                     /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"
     rep_origin      1283..1871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1772..1791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1933..2062
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2930..2939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2936..3646
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3081..3099)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3349..3368
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3671..4259
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3724..3744)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4142..4153)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4279..4755
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4478..4497)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4801..4941
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4801..4930
                     /label=AAV2 ITR
     rep_origin      5016..5471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5103..5122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5313..5334
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5488..5507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
       61 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
      121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      241 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      301 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      361 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      421 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      481 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      541 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      601 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      661 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      721 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      781 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      841 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      901 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      961 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1021 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1081 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1141 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1201 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1261 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1321 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1381 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1441 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1501 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1561 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1621 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1681 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1741 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1801 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1861 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1921 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
     1981 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     2041 aactccatca ctaggggttc ctgcggccgc acgcgttgca aagatggata aagttttaaa
     2101 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct
     2161 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2221 gggtcggcag cgctagatct aacatatcct ggtgtggagt aggggacgct gctctgacag
     2281 aggctcgggg gcctgagctg gctctgtgag ctggggagga ggcagacagc caggccttgt
     2341 ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc ttcatgccca
     2401 gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc ctgcttcccg
     2461 ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg ggcttgcatt
     2521 gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc ctaggggctg
     2581 cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc tgtgcccagg
     2641 aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga gaggagggct
     2701 gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc cccatagctg
     2761 ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag gggcacccgg
     2821 gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc gagcagagcc
     2881 agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggatccg ccaccatggt
     2941 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     3001 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     3061 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     3121 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     3181 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     3241 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     3301 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     3361 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     3421 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     3481 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     3541 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     3601 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagaat tctgtacaaa
     3661 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3721 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3781 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3841 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3901 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3961 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     4021 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     4081 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     4141 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     4201 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     4261 tcgataccgg ctcgagacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     4321 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     4381 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     4441 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     4501 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     4561 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     4621 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     4681 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     4741 cccttccctg tccttctcga gctgattttg taggtaacca cgtgcggacc gagcggccgc
     4801 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4861 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4921 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4981 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     5041 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     5101 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5161 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5221 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5281 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5341 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     5401 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5461 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5581 cgcttacaga caagctgtga ccgtct
//
