LOCUS       sequence-338414-        6444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Astrocyte AAV-mediated expression of jRCaMP1a, a red fluorescent 
            calcium sensor protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6444)
  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 6444)
  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 gfaABC1D NES-jRCaMP1a
FEATURES             Location/Qualifiers
     source          1..6444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   405..534
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(596..1184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(676..695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1355..2146,2147..2215))
                     /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"
     promoter        complement(2216..2320)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2388..2406
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2444..2466)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2566..2585
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2602..3057)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2739..2760)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2951..2970
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3132..3272
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3143..3272
                     /label=AAV2 ITR
     polyA_signal    3312..3788
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3570..3589
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     3855..3871
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3856..3872
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3875..4463
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3981..3992
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4390..4410
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5632..5688)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             complement(join(5716..5730,5731..5739))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(5743..5775)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(5779..5796)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5910..5929)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5912..6044
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
ORIGIN
        1 ggctgaatag agccttgttc tccaccgccg gtggcgccac acagcaaggg cagcccctag
       61 ggggctgtgc tgcttttatc ccaagatgcc aggctgtcag gctggggtgc aatgcaagcc
      121 ccctgctcaa tgggcttctc ggaaggcatt tgaggggggc tggggtgcgg cgggaagcag
      181 gcactgtgcc caccccgaac attgtgtctg tgccaaggtg agtcattcac tgggcatgaa
      241 gaggaggccc tggggggcgg ccagcccaat gctgccaggt ctgcttgcag acaaggcctg
      301 gctgtctgcc tcctccccag ctcacagagc cagctcaggc ccccgagcct ctgtcagagc
      361 agcgtcccct actccacacc aggatatgtt acgcgtgcgg ccgcaggaac ccctagtgat
      421 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
      481 cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggacatgt
      541 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      601 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      661 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      721 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      781 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      841 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      901 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      961 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1021 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1081 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1141 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1201 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1261 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1321 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1381 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1441 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1501 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1561 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1621 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1681 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1741 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1801 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1861 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1921 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1981 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2041 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2101 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2161 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2221 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2281 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2341 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     2401 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2461 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2521 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     2581 ttgtactgag agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa
     2641 atttttgtta aatcagctca ttttttaacc aatagaccga aatcggcaaa atcccttata
     2701 aatcaaaaga atagcccgag atagagttga gtgttgttcc agtttggaac aagagtccac
     2761 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2821 cactacgtga accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2881 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2941 cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg
     3001 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact
     3061 atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     3121 catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
     3181 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     3241 gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtggttacc
     3301 tacaaaatca gaaggacagg gaagggagca gtggttcacg cctgtaatcc cagcaatttg
     3361 ggaggccaag gtgggtagat cacctgagat taggagttgg agaccagcct ggccaatatg
     3421 gtgaaacccc gtctctacca aaaaaacaaa aattagctga gcctggtcat gcatgcctgg
     3481 aatcccaaca actcgggagg ctgaggcagg agaatcgctt gaacccagga ggcggagatt
     3541 gcagtgagcc aagattgtgc cactgcactc cagcttggtt cccaatagac cccgcaggcc
     3601 ctacaggttg tcttcccaac ttgccccttg ctccatacca cccccctcca ccccataata
     3661 ttatagaagg acacctagtc agacaaaatg atgcaactta attttattag gacaaggctg
     3721 gtgggcactg gagtggcaac ttccagggcc aggagaggca ctggggaggg gtcacaggga
     3781 tgccacccgt agatcatctc tagactcgag gtcgaggcca gagaggccat cgaatggcgc
     3841 gccgcgatcg cgtctcgagg tcgacggtat cgatgcgggg aggcggccca aagggagatc
     3901 cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc gcagagccgg cagcaggccg
     3961 cgggaaggaa ggtccgctgg attgagggcc gaagggacgt agcagaagga cgtcccgcgc
     4021 agaatccagg tggcaacaca ggcgagcagc caaggaaagg acgatgattt ccccgacaac
     4081 accacggaat tgtcagtgcc caacagccga gcccctgtcc agcagcgggc aaggcaggcg
     4141 gcgatgagtt ccgccgtggc aatagggagg gggaaagcga aagtcccgga aaggagctga
     4201 caggtggtgg caatgcccca accagtgggg gttgcgtcag caaacacagt gcacaccacg
     4261 ccacgttgcc tgacaacggg ccacaactcc tcataaagag acagcaacca ggatttatac
     4321 aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca
     4381 gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa
     4441 attttgtaat ccagaggttg attatcgata agcttactac ttcgctgtca tcatttgtac
     4501 aaactcttcg tagtttacct gaccatcccc atcgctgtct gcttccctga tcatttcatc
     4561 aacctcttca tctgttaact tctctccaag gtttgtcatc acgtggcgaa gctctgctgc
     4621 actgatgtag ccattgccat ccttatcaaa cacgccgaac gcttctctaa tttcttcttc
     4681 actgtctgtg tatttcattt ttcttgccat cataatcagg aactcaggga agtcgattgt
     4741 gccgtcaccg tcggcatcta cttcattgat catgtcctgc agctctgctt ctgtggggtt
     4801 ctgccccaga gaccgcatca ccgtccccag ctccttggtt gttattgtcc catccccgtc
     4861 cttgtcaaat agggagaaag cctctttaaa ttctgcgatc tgctcttcag tggtttggga
     4921 gtccgtaggc tcccaacctt tggttttctt ttgcattaca gcaccattgc tcgggaagtt
     4981 cacaccgcga acctgcacgt gatacaccag gcagccatcc tccagagacg tgtcttgcat
     5041 aacggtaatt acgccgccgt cttcgtaacg ggtcacacgt tcccaggtaa aaccttccgg
     5101 gaaggactgc ttaaagaaat ccgggatgcc tttcggatat ttgatgaagg tacgggagcc
     5161 atacataaag gacgtcgcca ggatgtcgaa cgcaaacggc agcggaccac cctcgatcac
     5221 tttgatacgc atagtctggg tgcccatata cgggttgcct tcgccttcac cagtgcattt
     5281 gaactggtgg ccgtttacgg agccttccag aaccactttc atacgcatgt tctccttgat
     5341 cagggagttc atgctccctc cggtaccgcc gccaccaccc aggcccacaa acttggcaac
     5401 tgcgtgttca cgctgtacga caaacatttc gttatcgctc tcctccaggc gctccagacg
     5461 gtggctgacg ctatgaatgg caggcatctt aatgttgccg acagtctttt tggagcggta
     5521 ggtggtcacg aaagaacagg acaggtgacc gccgccatca actttcagcg ccatgtgagt
     5581 gtaaccacgc agaccaccat ccgctgggta catcatttcg cagttgatcg ctgagctcag
     5641 ccgacctata gctctgactg cgtgacctgt cttattccac ttacgacgtg atgagtcgac
     5701 cattgttgcg agatccttat cgtcatcgtc gtacagatcc cgacccattt gctgtccacc
     5761 agtcatgcta gccataccat gatgatgatg atgatgagaa cctcctccag tcttgttaat
     5821 atcaagtcca gccaacttaa gagcaagctc gttctgcagc atggtggcgg gtggcggcga
     5881 attcagctgt aattgaactg ggagtggaca cctgtggaga gaaaggcaaa gtggatgtca
     5941 gtaagaccaa taggtgccta tcagaaacgc aagagtcttc tctgtctcga caagcccagt
     6001 ttctattggt ctccttaaac ctgtcttgta accttgatac ttacctgccc agtgcctcac
     6061 gaccaacttc tgcagcttaa gttcgagact gttgtgtcag aagcactgac tgcgttagca
     6121 atttaactgt gataaactac cgcaataaag gttgcgagca gcggaggtga tgcgtctcct
     6181 ctccaacctg ctctggctct gctcgctcct gggatgcgag ggctttatga aggagtgggc
     6241 tagactggcg atgcccgggt gcccctggca acaccccctg gcatagcctg agggggtggg
     6301 gttgggccgc agcccagcta tggggagagc tctcccctca cccatttgtg tccttggact
     6361 tctgggaagc agacagccct cctctccccc cctgccaccc actgtccatg cctgggcatc
     6421 ccctgatccc ctttcctggg caca
//
