LOCUS       sequence-369484-        6302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fluorescent voltage indicator Archon under the GFAP 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  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 6302)
  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-Archon-EGFP
FEATURES             Location/Qualifiers
     source          1..6302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      951..960
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1764..1766,1767..2480)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1764..1766,1767..2477)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1806..1827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2067..2086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2414..2435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2538..2579
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2597..3185
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2650..2670)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3068..3079)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3210..3227
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3250..3267)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3256..3480
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3565..3705
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3565..3694
                     /label=AAV2 ITR
     rep_origin      3780..4235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3867..3886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4077..4098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4252..4271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4371..4393
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4431..4449)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4517..4621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4622..4690,4691..5482)
                     /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      5653..6241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6142..6161
                     /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 tggtaccgag ctcggatccc gccaccatgg
      961 tttctatcgc actgcaggct ggttatgacc tcctcggaga tggtagacct gaaagtctgt
     1021 ggctcggaat tggtacactg ctgatgctga tcggaacctt ctacttcctg gttagagcgt
     1081 ggggagagac tgacaaggat gcacgggagt actacgcagt gacaatcctg gtcagcggta
     1141 ttgccagtgc agcttatctg tcaatgttct ttggtattgg cctcacagag gtgccagtag
     1201 gtggagagat gctgaacatc tactacgcaa gatacgcaca gtggctgttc actacaccac
     1261 tgttgctgct gcacctggca ctcctggcca aagttgatag agtgaccatc ggcactctcg
     1321 ttggcgtcga tgctctgatg attgtcacag gcctgatagg aacactgtca catactgcca
     1381 ttgccagata ctcttggtgg ctgttctcaa ctatctgtat gatcgtcgtt ctgtatgtcc
     1441 tggccacttc tctgcggtca gccgctaagg agagaggccc agaggttgcc tcaaccttca
     1501 acatcctcac cgcactggtt ctggtcctgt ggactgctta ccctatcatc tggattatcg
     1561 gcaccgaggg tgccggagtt gtgggcctgg gaatcgaaac tttgctgttc atggtgctgg
     1621 acgtcaccgc caaggtgggc ttcggtttta tcctgctcag gtcaagagcc attctgcaag
     1681 atacagaagc acctgaacct tcagctggag ctgataccgg tgccgccgac cgcccggtag
     1741 tagcagtgag caaggcggcc gccgtgagca agggcgagga gctgttcacc ggggtggtgc
     1801 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1861 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catttgcacc accggcaagc
     1921 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1981 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2041 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2101 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2161 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2221 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2281 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2341 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2401 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2461 tggacgagct gtacaagtaa gaattctgca gatatccagc acagtggcgg ccgctcgagt
     2521 ctagagggcc cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     2581 gtaccggtta atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2641 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2701 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2761 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2821 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2881 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2941 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3001 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3061 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3121 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3181 cccgcatcga aacccgctga tcagccggtc atcatcacca tcaccattga gtttaaaccc
     3241 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3301 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3361 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3421 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3481 cttctgaggc ggaaagaacc agatcctctc ttaaggtagc atcgagattt aaattaggga
     3541 taacagggta atggcgcggg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3601 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3661 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3721 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3781 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3841 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3901 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3961 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4021 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4081 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     4141 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4201 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4261 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4321 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4381 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4441 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4501 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4561 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4621 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4681 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4741 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4801 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4861 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4921 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4981 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5041 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5101 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5161 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5221 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5281 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5341 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5401 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5461 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5521 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5581 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5761 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5881 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5941 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6001 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6061 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6121 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6181 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6241 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6301 gt
//
