LOCUS       sequence-115796-        4123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genetically encodable fluorescent calcium indicator protein to 
            monitor neuronal activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4123)
  AUTHORS   Badura A, Sun XR, Giovannucci A, Lynch LA, Wang SS
  TITLE     Fast calcium sensor proteins for monitoring neural activity.
  JOURNAL   Neurophotonics. 2014 Oct;1(2):025008.
  PUBMED    25558464
REFERENCE   2  (bases 1 to 4123)
  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: 
            "Neurophotonics."; date: "2014-10"; volume: "1(2)"; pages: "025008"
COMMENT     Sequence Label: Fast-GCaMP6f-RS06
FEATURES             Location/Qualifiers
     source          1..4123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(100..108,109..126,127..129,130..162,163..165,166..189,
                     190..216,217..273,274..279,280..549,550..567,568..1002,
                     1003..1008,1009..1452)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMQARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      196..205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     486..507
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(613..634)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(874..893)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1524..1542)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1538..1585
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1682..2137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1769..1788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1979..2000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2163..2267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2268..2336,2337..3128)
                     /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(2486..2505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3299..3887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3788..3807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4041..4058
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgggttctca tcatcatcat
      121 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      181 gacgataagg atctcgccac catggtcgac tcatcacgtc gtaagtggaa taagacaggt
      241 cacgcagtca gagctatagg tcggctgagc tcactcgaga acgtctatat caaggccgac
      301 aagcagaaga acggcatcaa ggcgaacttc aagatccgcc acaacatcga ggacggcggc
      361 gtgcagctcg cctaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      421 cccgacaacc actacctgag cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc
      481 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      541 ctgtacaagg gcggtaccgg agggagcatg gtgagcaagg gcgaggagct gttcaccggg
      601 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      661 ggcgagggtg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      721 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      781 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      841 ggctacatcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      901 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      961 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acctgccgga ccaactgact
     1021 gaagagcaga tcgcagaatt taaagaggaa ttctccctat ttgacaagga cggggatggg
     1081 acaataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa
     1141 gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc
     1201 cctgagttcc tgacaatgca ggcaagaaaa atgaaataca gggacacgga agaagaaatt
     1261 agagaagcgt tcggtgtgtt tgataaggat ggcaatggct acatcagtgc agcagagctt
     1321 cgccacgtga tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc
     1381 agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg
     1441 acagcgaagt aaggtgtaca aagatatcaa gcttgatccg gctgctaaca aagcccgaaa
     1501 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1561 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat
     1621 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1681 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1741 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1801 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1861 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1921 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1981 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2041 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2101 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     2161 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2221 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2281 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2341 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2401 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2461 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2521 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2581 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2641 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2701 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2761 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2821 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2881 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2941 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3001 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3061 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3121 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3181 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3241 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3301 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3361 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3421 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3481 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3541 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3601 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3661 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3721 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3781 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3841 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3901 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3961 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4021 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4081 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cag
//
