LOCUS       sequence-360464-        6374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFAP-ER-GCaMP6f AAV: An AAV5 constructed for ER-GCaMP6f with 
            astrocyte specific GFAP promotor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  AUTHORS   Aryal SP, Xia M, Adindu E, Davis C, Ortinski PI, Richards CI
  TITLE     ER-GCaMP6f: An Endoplasmic Reticulum-Targeted Genetic Probe to 
            Measure Calcium Activity in Astrocytic Processes.
  JOURNAL   Anal Chem. 2022 Jan 21. doi: 10.1021/acs.analchem.1c04321.
  PUBMED    35061939
REFERENCE   2  (bases 1 to 6374)
  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: "Anal Chem. 
            2022 Jan 21. doi: 10.1021/acs.analchem.1c04321."
COMMENT     Sequence Label: ER-GCaMP6f-AAV
FEATURES             Location/Qualifiers
     source          1..6374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   204..344
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   204..333
                     /label=AAV2 ITR
     rep_origin      419..874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(506..525)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     716..737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(891..910)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1010..1032
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1070..1088)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1156..1260
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1261..1329,1330..2121)
                     /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      2292..2880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2781..2800
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2942..3071
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      4056..4065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4077..4133
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     4346..4367
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4473..4494)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4734..4753)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5448..6036
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5501..5521)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5919..5930)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(6056..6374,1..158)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6255..6274)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
       61 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
      121 gattacaggc gtgaaccact gctcccttcc ctgtccttct cgagctgatt ttgtaggtaa
      181 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
      241 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
      301 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
      361 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
      421 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      481 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      541 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      601 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
      661 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
      721 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
      781 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
      841 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
      901 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
      961 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     1021 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     1081 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     1141 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1201 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1261 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1321 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1381 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1441 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1501 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1561 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1621 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1681 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1741 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1801 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1861 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1921 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1981 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2041 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2101 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2161 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2221 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2281 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2341 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2401 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2461 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2521 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2581 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2641 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2701 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2761 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2821 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2881 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2941 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     3001 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     3061 taggggttcc tgcggccgca cgcgttgcaa agatggataa agttttaaac agagaggaat
     3121 ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt
     3181 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcagc
     3241 gctagatcta acatatcctg gtgtggagta ggggacgctg ctctgacaga ggctcggggg
     3301 cctgagctgg ctctgtgagc tggggaggag gcagacagcc aggccttgtc tgcaagcaga
     3361 cctggcagca ttgggctggc cgccccccag ggcctcctct tcatgcccag tgaatgactc
     3421 accttggcac agacacaatg ttcggggtgg gcacagtgcc tgcttcccgc cgcaccccag
     3481 cccccctcaa atgccttccg agaagcccat tgagcagggg gcttgcattg caccccagcc
     3541 tgacagcctg gcatcttggg ataaaagcag cacagccccc taggggctgc ccttgctgtg
     3601 tggcgccacc ggcggtggag aacaaggctc tattcagcct gtgcccagga aaggggatca
     3661 ggggatgccc aggcatggac agtgggtggc agggggggag aggagggctg tctgcttccc
     3721 agaagtccaa ggacacaaat gggtgagggg agagctctcc ccatagctgg gctgcggccc
     3781 aaccccaccc cctcaggcta tgccaggggg tgttgccagg ggcacccggg catcgccagt
     3841 ctagcccact ccttcataaa gccctcgcat cccaggagcg agcagagcca gagcaggttg
     3901 gagaggagac gcatcacctc cgctgctcgc ggggatctcg agccaccatg catccggata
     3961 tggaccctgt ggtcgtgctg gggctgtgcc tgagctgtct gctgctgctg agcctgtgga
     4021 agcagtccta cggaggcggg ttggatccac cggtcgccac catggtcgac tcatcacgtc
     4081 gtaagtggaa taagacaggt cacgcagtca gagctatagg tcggctgagc tcactcgaga
     4141 acgtctatat caaggccgac aagcagaaga acggcatcaa ggcgaacttc aagatccgcc
     4201 acaacatcga ggacggcggc gtgcagctcg cctaccacta ccagcagaac acccccatcg
     4261 gcgacggccc cgtgctgctg cccgacaacc actacctgag cgtgcagtcc aaactttcga
     4321 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     4381 tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg gtgagcaagg
     4441 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     4501 gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc aagctgaccc
     4561 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     4621 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     4681 tcaagtccgc catgcccgaa ggctacatcc aggagcgcac catcttcttc aaggacgacg
     4741 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     4801 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     4861 acctgccgga ccaactgact gaagagcaga tcgcagaatt taaagaggaa ttctccctat
     4921 ttgacaagga cggggatggg acaataacaa ccaaggagct ggggacggtg atgcggtctc
     4981 tggggcagaa ccccacagaa gcagagctgc aggacatgat caatgaagta gatgccgacg
     5041 gtgacggcac aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa atgaaataca
     5101 gggacacgga agaagaaatt agagaagcgt tcggtgtgtt tgataaggat ggcaatggct
     5161 acatcagtgc agcagagctt cgccacgtga tgacaaacct tggagagaag ttaacagatg
     5221 aagaggttga tgaaatgatc agggaagcag acatcgatgg ggatggtcag gtaaactacg
     5281 aagagtttgt acaaatgatg acagcgaagc tttaaagcgg ccgcgactct agatcataat
     5341 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     5401 gaacctgaaa cataaaatga atgcaattct gtacaaagct tatcgataat caacctctgg
     5461 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5521 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5581 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5641 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5701 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5761 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5821 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     5881 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5941 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6001 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccggctc gagacgggtg
     6061 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     6121 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     6181 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     6241 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     6301 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     6361 attccaggca tgca
//
