LOCUS       sequence-369489-        6218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fluorescent voltage indicator Arclight under the GFAP 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Armbruster M, Naskar S, Garcia JP, Sommer M, Kim E, Adam Y, Haydon 
            PG, Boyden ES, Cohen AE, Dulla CG
  TITLE     Neuronal activity drives pathway-specific depolarization of 
            peripheral astrocyte processes.
  JOURNAL   Nat Neurosci. 2022 May;25(5):607-616. doi: 
            10.1038/s41593-022-01049-x. Epub 2022 Apr 28.
  PUBMED    35484406
REFERENCE   2  (bases 1 to 6218)
  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"; doi: 
            "10.1038/s41593-022-01049-x"; journalName: "Nat Neurosci"; date: 
            "2022-05"; volume: "25"; issue: "5"; pages: "607-616"
COMMENT     Sequence Label: GFAP-Arclight
FEATURES             Location/Qualifiers
     source          1..6218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1667..2383
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTADGITHGMDELYK"
     primer_bind     complement(1697..1725)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2320..2340
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2454..2495
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2513..3101
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2566..2586)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2984..2995)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3126..3143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3166..3183)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3172..3396
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3481..3621
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3481..3610
                     /label=AAV2 ITR
     rep_origin      3696..4151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3783..3802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3993..4014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4168..4187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4287..4309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4347..4365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4433..4537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4538..4606,4607..5398)
                     /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      5569..6157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6058..6077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcaattc agtcgataac tataacggtc ctaaggtagc gatttaaata
      181 cgcgctctct taaggtagcc ccgggacgcg tcaattgaga acatatcctg gtgtggagta
      241 ggggacgctg ctctgacaga ggctcggggg cctgagctgg ctctgtgagc tggggaggag
      301 gcagacagcc aggccttgtc tgcaagcaga cctggcagca ttgggctggc cgccccccag
      361 ggcctcctct tcatgcccag tgaatgactc accttggcac agacacaatg ttcggggtgg
      421 gcacagtgcc tgcttcccgc cgcaccccag cccccctcaa atgccttccg agaagcccat
      481 tgagcagggg gcttgcattg caccccagcc tgacagcctg gcatcttggg ataaaagcag
      541 cacagccccc taggggctgc ccttgctgtg tggcgccacc ggcggtggag aacaaggctc
      601 tattcagcct gtgcccagga aaggggatca ggggatgccc aggcatggac agtgggtggc
      661 agggggggag aggagggctg tctgcttccc agaagtccaa ggacacaaat gggtgagggg
      721 actggagggg agctctcccc atagctgggc tgcggcccaa ccccaccccc tcaggctatg
      781 ccagggggtg ttgccagggg cacccgggca tcgccagtct agcccactcc ttcataaagc
      841 cctcgcatcc caggagcgag cagagccaga gcaggttgga gaggagacgc atcacctccg
      901 ctgctcgccg ctagcgttta aacttaagct tatggaggga ttcgacggtt cagattttag
      961 tcctccagct gatttagttg gcgttgacgg tgcagtcatg cggaacgtcg ttgacgtcac
     1021 gataaatggt gacgtcactg ctccgccgaa agctgcgcca agaaaaagtg aatcggtaaa
     1081 gaaagttcat tggaatgatg tagaccaagg accgagtgaa aaaccagaga caagacagga
     1141 ggaacgaata gatatacccg agatttcagg tctatggtgg ggcgagaatg aacatggagt
     1201 ggacgatggg agaatggaga tacctactac tggtgtaggt cgcgtccagt ttcgtgtccg
     1261 agcagtgatt gatcatctag ggatgcgagt ctttggagtc ttcctaattt tcttggacat
     1321 catcctcatg atcattgatc tcagtcttcc aggaaaaagt gaatcttcac aatcctttta
     1381 tgacgggatg gctttggctc tttcttgtta tttcatgctg gatttaggat taaggatatt
     1441 tgcctacggg cccaagaatt tcttcaccaa cccctgggag gttgctgatg gtttgattat
     1501 cgttgtcaca ttcgtcgtca cgatatttta cactgtgtta gatgaatacg tgcaagaaac
     1561 aggagccgat ggtttggggc aattggttgt gttggcccgt ttgctgcgtg tggttcgatt
     1621 agcaagaata ttttattccc accaacaaat gaaggctggg gatcccatga gtaaaggaga
     1681 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
     1741 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
     1801 atttatttgc actactggaa aactacctgt tccatggcca acacttgtca ctactttaac
     1861 ttatggtgtt caatgctttt caagataccc agatcatatg aaacggcatg actttttcaa
     1921 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
     1981 ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt
     2041 aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta
     2101 taacgatcac caggtgtaca tcatggcaga caaacaaaag aatggaatca aagctaactt
     2161 caaaattaga cacaacattg aagatggagg cgttcaacta gcagaccatt atcaacaaaa
     2221 tactccaatt ggcgatgggc ccgtcctttt accagacaac cattacctgt ttacaacttc
     2281 tactctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac
     2341 agctgatggg attacacatg gcatggatga actatacaaa tgatctagaa ctatagtgag
     2401 tcgtattaca tccagcacag tggcggccgc tcgagtctag agggcccttc gaaggtaagc
     2461 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttaatcg ataatcaacc
     2521 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2581 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2641 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2701 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2761 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2821 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2881 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     2941 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3001 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3061 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgaaacc cgctgatcag
     3121 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     3181 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3241 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3301 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3361 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagat
     3421 cctctcttaa ggtagcatcg agatttaaat tagggataac agggtaatgg cgcgggccgc
     3481 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3541 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3601 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3661 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3901 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4021 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4141 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4201 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4261 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4321 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4381 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4441 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4501 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4561 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4621 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4681 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4741 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4801 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4861 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4921 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4981 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5041 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5101 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5161 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5221 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5281 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5341 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5401 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5461 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5521 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5581 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5641 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5701 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5761 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5821 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5881 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5941 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6001 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6061 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6181 acggttcctg gccttttgct ggccttttgc tcacatgt
//
