LOCUS       sequence-183009-        5308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ultrasensitive protein calcium sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5308)
  AUTHORS   Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, 
            Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, 
            Kim DS
  TITLE     Ultrasensitive fluorescent proteins for imaging neuronal activity.
  JOURNAL   Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
  PUBMED    23868258
REFERENCE   2  (bases 1 to 5308)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2013-07-18- 18"; volume: "499"; issue: "7458"; 
            pages: "295-300"
COMMENT     Sequence Label: pGP-CMV-GCaMP6f
FEATURES             Location/Qualifiers
     source          1..5308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        137..328
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     285..305
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(330..434)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      461..916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(548..567)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     758..779
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    923..1044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(934..953)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     988..1007
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1166..1609,1610..1615,1616..2050,2051..2068,
                     2069..2338,2339..2344,2345..2401,2402..2428,2429..2452,
                     2453..2455,2456..2488,2489..2491,2492..2509,2510..2518))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1725..1744
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1984..2005
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2111..2132)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2413..2422
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2515..2524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2572..2775)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2601..2621)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2776..3079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3254..3842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3334..3353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4171..4218
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4243..4262
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4450..5244)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4450..5244)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4561..4580)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5171..5190
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5279..5308,1..328))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5293..5308,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
       61 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
      121 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
      241 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
      301 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt
      361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
      421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
      481 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      541 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      601 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      661 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      721 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      781 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      841 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      901 ttaacgctta caatttacgc cttaagatac attgatgagt ttggacaaac cacaactaga
      961 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1021 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1081 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     1141 gattatgatc tagagtcgcg gccgctcact tcgctgtcat catttgtaca aactcttcgt
     1201 agtttacctg accatcccca tcgatgtctg cttccctgat catttcatca acctcttcat
     1261 ctgttaactt ctctccaagg tttgtcatca cgtggcgaag ctctgctgca ctgatgtagc
     1321 cattgccatc cttatcaaac acaccgaacg cttctctaat ttcttcttcc gtgtccctgt
     1381 atttcatttt tcttgccatc attgtcagga actcagggaa gtcgattgtg ccgtcaccgt
     1441 cggcatctac ttcattgatc atgtcctgca gctctgcttc tgtggggttc tgccccagag
     1501 accgcatcac cgtccccagc tccttggttg ttattgtccc atccccgtcc ttgtcaaata
     1561 gggagaattc ctctttaaat tctgcgatct gctcttcagt cagttggtcc ggcaggttgt
     1621 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     1681 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     1741 cgtccttgaa gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga
     1801 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     1861 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     1921 tcagcttgcc gtaggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt
     1981 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     2041 tgctcaccat gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc
     2101 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg
     2161 aaagtttgga ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2221 tgggggtgtt ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc
     2281 ggatcttgaa gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacgttct
     2341 cgagtgagct cagccgacct atagctctga ctgcgtgacc tgtcttattc cacttacgac
     2401 gtgatgagtc gaccatggtg gcgagatcct tatcgtcatc gtcgtacaga tcccgaccca
     2461 tttgctgtcc accagtcatg ctagccatac catgatgatg atgatgatga gaacccatgg
     2521 tggcgagatc tgagtccggt agcgctagcg gatctgacgg ttcactaaac cagctctgct
     2581 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     2641 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2701 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2761 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2821 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2881 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2941 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     3001 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     3061 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     3121 tgattactat taataactaa tgcatggcgg taatacggtt atccacagaa tcaggggata
     3181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3961 aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg
     4021 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg
     4081 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt
     4141 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat
     4201 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt
     4261 cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga
     4321 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     4381 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     4441 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     4501 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     4561 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     4621 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     4681 atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc
     4741 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     4801 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     4861 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     4921 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     4981 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     5041 cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac
     5101 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     5161 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     5221 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt
     5281 tgcaaaagcc taggcctcca aaaaagcc
//
