LOCUS       sequence-338268-        7841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Astrocyte AAV-mediated expression of bright ultrasensitive protein 
            calcium sensor under the GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7841)
  AUTHORS   Lohr C, Beiersdorfer A, Fischer T, Hirnet D, Rotermund N, Sauer J, 
            Schulz K, Gee CE
  TITLE     Using Genetically Encoded Calcium Indicators to Study Astrocyte 
            Physiology: A Field Guide.
  JOURNAL   Front Cell Neurosci. 2021 Jun 11;15:690147. doi: 
            10.3389/fncel.2021.690147. eCollection 2021.
  PUBMED    34177468
REFERENCE   2  (bases 1 to 7841)
  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: "Front Cell 
            Neurosci."; date: "2021-06-11"; volume: "15:690147. doi"; pages: " 
            10.3389/fncel.2021.690147. eCollection 2021"
COMMENT     Sequence Label: pAAV GFAP jGCaMP7b
FEATURES             Location/Qualifiers
     source          1..7841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   11..140
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     misc_feature    156..273
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      2552..2561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2558..2566,2567..2584,2585..2587,2588..2620,2621..2623,
                     2624..2647,2648..2674,2675..2731,2732..2737,2738..3007,
                     3008..3025,3026..3460,3461..3466,3467..3910)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNPPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMYARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      2654..2663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2944..2965
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3071..3092)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3332..3351)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3923..4511
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3976..3996)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4394..4405)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4514..4530)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4515..4531)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4598..5074
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4797..4816)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5114..5254
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5114..5243
                     /label=AAV2 ITR
     rep_origin      5329..5784
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5416..5435)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5626..5647
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5801..5820)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5920..5942
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5980..5998)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6066..6170
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6171..6239,6240..7031)
                     /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      7202..7790
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7691..7710
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
       61 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
      121 ctccatcact aggggttcct gcggccgcac gcgtttttta aaagaaaagg ggggattggg
      181 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
      241 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
      301 ccagtttggt taattaaccc acctccctct ctgtgctggg actcacagag ggagacctca
      361 ggaggcagtc tgtccatcac atgtccaaat gcagagcata ccctgggctg ggcgcagtgg
      421 cgcacaactg taattccagc actttgggag gctgatgtgg aaggatcact tgagcccaga
      481 agttctagac cagcctgggc aacatggcaa gaccctatct ctacaaaaaa agttaaaaaa
      541 tcagccacgt gtggtgacac acacctgtag tcccagctat tcaggaggct gaggtgaggg
      601 gatcacttaa ggctgggagg ttgaggctgc agtgagtcgt ggttgcgcca ctgcactcca
      661 gcctgggcaa cagtgagacc ctgtctcaaa agacaaaaaa aaaaaaaaaa aaaaagaaca
      721 tatcctggtg tggagtaggg gacgctgctc tgacagaggc tcgggggcct gagctggctc
      781 tgtgagctgg ggaggaggca gacagccagg ccttgtctgc aagcagacct ggcagcattg
      841 ggctggccgc cccccagggc ctcctcttca tgcccagtga atgactcacc ttggcacaga
      901 cacaatgttc ggggtgggca cagtgcctgc ttcccgccgc accccagccc ccctcaaatg
      961 ccttccgaga agcccattga gcagggggct tgcattgcac cccagcctga cagcctggca
     1021 tcttgggata aaagcagcac agccccctag gggctgccct tgctgtgtgg cgccaccggc
     1081 ggtggagaac aaggctctat tcagcctgtg cccaggaaag gggatcaggg gatgcccagg
     1141 catggacagt gggtggcagg gggggagagg agggctgtct gcttcccaga agtccaagga
     1201 cacaaatggg tgaggggact gggcagggtt ctgaccctgt gggaccagag tggagggcgt
     1261 agatggacct gaagtctcca gggacaacag ggcccaggtc tcaggctcct agttgggccc
     1321 agtggctcca gcgtttccaa acccatccat ccccagaggt tcttcccatc tctccaggct
     1381 gatgtgtggg aactcgagga aataaatctc cagtgggaga cggaggggtg gccagggaaa
     1441 cggggcgctg caggaataaa gacgagccag cacagccagc tcatgtgtaa cggctttgtg
     1501 gagctgtcaa ggcctggtct ctgggagaga ggcacaggga ggccagacaa ggaaggggtg
     1561 acctggaggg acagatccag gggctaaagt cctgataagg caagagagtg ccggccccct
     1621 cttgccctat caggacctcc actgccacat agaggccatg attgaccctt agacaaaggg
     1681 ctggtgtcca atcccagccc ccagccccag aactccaggg aatgaatggg cagagagcag
     1741 gaatgtggga catctgtgtt caagggaagg actccaggag tctgctggga atgaggccta
     1801 gtaggaaatg aggtggccct tgagggtaca gaacaggttc attcttcgcc aaattcccag
     1861 caccttgcag gcacttacag ctgagtgaga taatgcctgg gttatgaaat caaaaagttg
     1921 gaaagcaggt cagaggtcat ctggtacagc ccttccttcc cttttttttt tttttttttt
     1981 gtgagacaag gtctctctct gttgcccagg ctggagtggc gcaaacacag ctcactgcag
     2041 cctcaaccta ctgggctcaa gcaatcctcc agcctcagcc tcccaaagtg ctgggattac
     2101 aagcatgagc caccccactc agccctttcc ttccttttta attgatgcat aataattgta
     2161 agtattcatc atggtccaac caaccctttc ttgacccacc ttcctagaga gagggtcctc
     2221 ttgcttcagc ggtcagggcc ccagacccat ggtctggctc caggtaccac ctgcctcatg
     2281 caggagttgg cgtgcccagg aagctctgcc tctgggcaca gtgacctcag tggggtgagg
     2341 ggagctctcc ccatagctgg gctgcggccc aaccccaccc cctcaggcta tgccaggggg
     2401 tgttgccagg ggcacccggg catcgccagt ctagcccact ccttcataaa gccctcgcat
     2461 cccaggagcg agcagagcca gagcaggttg gagaggagac gcatcacctc cgctgctcgc
     2521 cggaattcaa gctgctagca aggatccacc cgccaccatg ggttctcatc atcatcatca
     2581 tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga
     2641 cgataaggat ctcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca
     2701 cgcagtcaga gctataggtc ggctgagctc actcgagaac gtctatatca aggccgacaa
     2761 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt
     2821 gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2881 cgacaaccac tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga
     2941 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3001 gtacaagggc ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt
     3061 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3121 cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3181 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     3241 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3301 ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3361 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag cttaagggca tcgacttcaa
     3421 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ccacctgacc aactgactga
     3481 agagcagatc gcagaattta aagagctttt ctccctattt gacaaggacg gggatgggac
     3541 aataacaacc aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc
     3601 agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc
     3661 tgagttcctg acaatgtacg caagaaaaat gaaatacagg gacacggaag aagaaattag
     3721 agaagcgttc ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg
     3781 ccacgtgatg acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag
     3841 ggaagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac
     3901 agcgaagtga aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3961 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4021 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4081 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4141 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4201 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4261 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4321 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4381 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4441 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4501 cgcctccccg catcgatacc gtcgacctcg agacgcgatc gcggcgcgcc attcgatggc
     4561 ctctctggcc tcgacctcga gtctagagat gatctacggg tggcatccct gtgacccctc
     4621 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     4681 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     4741 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     4801 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     4861 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     4921 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     4981 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     5041 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     5101 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5161 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5221 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     5281 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtaa gcgccctgta
     5341 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5401 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5461 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5521 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5581 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5641 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggatttttc
     5701 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5761 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     5821 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5881 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5941 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6001 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6061 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6121 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6181 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6241 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6301 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6361 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6421 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6481 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6541 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6601 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6661 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6721 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6781 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6841 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6901 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6961 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7021 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7081 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7141 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7201 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7261 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7321 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7381 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7441 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7501 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7561 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7621 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7681 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7741 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7801 acgcggcctt tttacggttc ctggcctttt gctggccttt t
//
