LOCUS       sequence-229529-        6039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Improved genetic encoded calcium sensor based on GCaMP6m.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  AUTHORS   Yang Y, Liu N, He Y, Liu Y, Ge L, Zou L, Song S, Xiong W, Liu X
  TITLE     Improved calcium sensor GCaMP-X overcomes the calcium channel 
            perturbations induced by the calmodulin in GCaMP.
  JOURNAL   Nat Commun. 2018 Apr 17;9(1):1504. doi: 10.1038/s41467-018-03719-6.
  PUBMED    29666364
REFERENCE   2  (bases 1 to 6039)
  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: "Nat 
            Commun."; date: "2018-04-17"; volume: "9(1):1504. doi"; pages: " 
            10.1038/s41467-018-03719-6"
COMMENT     Sequence Label: AAV-syn-GCaMP6m-XC
FEATURES             Location/Qualifiers
     source          1..6039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..579
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(716..724,725..742,743..745,746..778,779..781,782..805,
                     806..832,833..889,890..895,896..1165,1166..1183,1184..1618,
                     1619..1624,1625..2065)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      812..821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1102..1123
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1229..1250)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1490..1509)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2123..2711
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2176..2196)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2594..2605)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2743..3219
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2942..2961)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3259..3399
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3259..3388
                     /label=AAV2 ITR
     rep_origin      3474..3929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3771..3792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3946..3965)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4065..4087
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4125..4143)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4211..4315
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4316..4384,4385..5176)
                     /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(4534..4553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5347..5935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5836..5855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5997..6039,1..87)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
       61 tggccaactc catcactagg ggttcctgcg gccgcacgcg tgtgtctaga ctgcagaggg
      121 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      181 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      241 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      301 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      361 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      421 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      481 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      541 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaaggta ccggatccgc
      601 caccatgggc atggacgagc tgtacaaggg taccgctgca accaaaattc aggctgcctt
      661 ccgtggacac ataacaagga aaaagctcaa agacgagaag aagggtgctg aattcatggg
      721 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      781 ggatctgtac gacgatgacg ataaggatct cgccaccatg gtcgactcat cacgtcgtaa
      841 gtggaataag acaggtcacg cagtcagagc tataggtcgg ctgagctcac tcgagaacgt
      901 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
      961 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1021 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     1081 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1141 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga
     1201 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1261 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa
     1321 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1381 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1441 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     1501 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1561 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct
     1621 gccggaccaa ctgactgaag agcagatcgc agaatttaaa gaggctttct ccctatttga
     1681 caaggacggg gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctggg
     1741 gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga
     1801 cggcacaatc gacttccctg agttcctgac aatgatggca agaaaaggga gctacaggga
     1861 cacggaagaa gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat
     1921 cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga
     1981 ggttgatgaa atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga
     2041 gtttgtacaa atgatgacag cgaagcttgc tcttaagttg gctggacttg atattggttc
     2101 ttaagatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2161 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2221 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2281 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2341 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2401 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2461 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2521 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     2581 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2641 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2701 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     2761 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2821 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2881 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2941 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3001 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3061 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3121 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3181 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3241 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3301 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3361 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3421 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3481 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3541 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3601 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3661 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3721 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3781 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3841 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3901 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3961 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4021 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4081 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4141 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4201 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4261 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4321 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4381 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4441 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4501 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4561 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4621 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4681 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4741 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4801 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4861 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4921 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4981 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5041 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5101 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5161 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg
     6001 caggcagctg cgcgctcgct cgctcactga ggccgcccg
//
