LOCUS       sequence-376574-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Improved genetic encoded calcium sensor based on GCaMP7b..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  AUTHORS   Geng J, Tang Y, Yu Z, Gao Y, Li W, Lu Y, Wang B, Zhou H, Li P, Liu 
            N, Wang P, Fan Y, Yang Y, Guo ZV, Liu X
  TITLE     Chronic Ca(2+) imaging of cortical neurons with long-term expression
            of GCaMP-X.
  JOURNAL   eLife. 2022 Oct 5;11:e76691. doi: 10.7554/eLife.76691.
  PUBMED    36196992
REFERENCE   2  (bases 1 to 6073)
  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: "eLife."; 
            date: "2022-10-5"; volume: "11:e76691. doi"; pages: " 
            10.7554/eLife.76691"
COMMENT     Sequence Label: AAV-hSyn-jGCaMP7b-XC
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        180..627
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      675..684
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(792..800,801..818,819..821,822..854,855..857,858..881,
                     882..908,909..965,966..971,972..1241,1242..1259,1260..1694,
                     1695..1700,1701..2141)
                     /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      888..897
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1178..1199
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1305..1326)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1566..1585)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2205..2793
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2258..2278)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2676..2687)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2825..3301
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3024..3043)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3341..3481
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3341..3470
                     /label=AAV2 ITR
     rep_origin      3556..4011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3643..3662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3853..3874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4028..4047)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4147..4169
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4207..4225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4293..4397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4398..4466,4467..5258)
                     /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"
     primer_bind     complement(4616..4635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5429..6017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5918..5937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatcctccg ctagcgctac
      661 cggactcaga tctcgccacc atgggcatgg acgagctgta caagggtacc gctgcaacca
      721 aaattcaggc tgccttccgt ggacacataa caaggaaaaa gctcaaagac gagaagaagg
      781 gtgctgaatt catgggttct catcatcatc atcatcatgg tatggctagc atgactggtg
      841 gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatctcgcc accatggtcg
      901 actcatcacg tcgtaagtgg aataagacag gtcacgcagt cagagctata ggtcggctga
      961 gctcactcga gaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact
     1021 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga
     1081 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt
     1141 ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1201 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca
     1261 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1321 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg
     1381 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1441 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1501 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct
     1561 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1621 tgaaccgcat cgagcttaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1681 agctggagta caacccacct gaccaactga ctgaagagca gatcgcagaa tttaaagagc
     1741 ttttctccct atttgacaag gacggggatg ggacaataac aaccaaggag ctggggacgg
     1801 tgatgcggtc tctggggcag aaccccacag aagcagagct gcaggacatg atcaatgaag
     1861 tagatgccga cggtgacggc acaatcgact tccctgagtt cctgacaatg tacgcaagaa
     1921 aaatgaaata cagggacacg gaagaagaaa ttagagaagc gttcggtgtg tttgataagg
     1981 atggcaatgg ctacatcagt gcagcagagc ttcgccacgt gatgacaaac cttggagaga
     2041 agttaacaga tgaagaggtt gatgaaatga tcagggaagc agacatcgat ggggatggtc
     2101 aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gcttgctctt aagttggctg
     2161 gacttgatat tggttcttaa gaattcgata tcaagcttat cgataatcaa cctctggatt
     2221 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2281 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2341 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2401 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2461 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2521 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2581 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     2641 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2701 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2761 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     2821 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     2881 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     2941 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     3001 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     3061 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     3121 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     3181 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     3241 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     3301 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     3361 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3421 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3481 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3541 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3601 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3661 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3721 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3781 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3841 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     3901 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     3961 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4021 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4081 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4141 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4201 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4261 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4321 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4381 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4441 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4501 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4561 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4621 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4681 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4741 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4801 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4861 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4921 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4981 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5041 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5101 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5161 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5221 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5281 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5341 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5401 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5461 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5521 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5581 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5641 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5701 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5761 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5821 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5881 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5941 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6001 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6061 ggccttttgc tca
//
