LOCUS       sequence-218119-        7125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically-encoded ratiometric bioluminescent Ca2+ indicator based 
            on GCaMP6s.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7125)
  AUTHORS   Qian Y, Rancic V, Wu J, Ballanyi K, Campbell RE
  TITLE     A bioluminescent Ca2+ indicator based on a topological variant of 
            GCaMP6s.
  JOURNAL   Chembiochem. 2018 Jun 22. doi: 10.1002/cbic.201800255.
  PUBMED    29934970
REFERENCE   2  (bases 1 to 7125)
  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: 
            "Chembiochem. 2018 Jun 22. doi: 10.1002/cbic.201800255."
COMMENT     Sequence Label: pcDNA-LUCI-GECO1
FEATURES             Location/Qualifiers
     source          1..7125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      672..681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             690..1202
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(1248..1269)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1509..1528)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2112..2168
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     2381..2402
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2482..2499)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2488..2712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2758..3186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2845..2864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3055..3076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3195..3215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3200..3529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3380..3515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3442..3461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3596..4390
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3650..3669)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4260..4279
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4564..4685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4601..4620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4655..4674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4734..4750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4734..4750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4747..4769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4758..4774
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4782..4788,4789..4806,4807..4812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4827..4848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4965..4982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5136..5724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5216..5235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5895..6686,6687..6755))
                     /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     6518..6537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6756..6860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6935..6954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgtgc cgccaccatg gcctcgagaa tggtcttcac actcgaagat ttcgttgggg
      721 actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca
      781 gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg
      841 aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc
      901 aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta
      961 aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact
     1021 atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag
     1081 ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc
     1141 tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg
     1201 cgcggaagag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1261 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct
     1321 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1381 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     1441 agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct
     1501 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1561 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1621 acaagctgga gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatttaaag
     1681 aggctttctc cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga
     1741 cggtgatgcg gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg
     1801 aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa
     1861 gaaaaatgaa atacagggac acggaagaag aaattagaga agcgttcggt gtgtttgata
     1921 aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag
     1981 agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc gatggggatg
     2041 gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaagggtggc ggaggttctg
     2101 tcgactcatc acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc
     2161 tgagctcact cgagaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga
     2221 acttccacat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc
     2281 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc
     2341 agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2401 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagcg gccgcgacaa
     2461 gcttggtaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2521 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2581 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2641 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2701 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2941 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3061 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3181 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3541 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3601 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3661 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3721 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3781 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3841 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3901 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3961 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4021 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4081 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4141 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4201 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4261 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4321 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4381 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4441 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4501 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4561 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4621 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4681 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4741 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4801 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4861 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4921 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4981 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5221 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5281 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5341 tataggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5401 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5461 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5521 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5581 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5641 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5701 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5761 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5821 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5881 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5941 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6001 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6061 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6121 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6181 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6241 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6301 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6361 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6421 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6481 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6541 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6601 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6661 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6721 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6781 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6841 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6901 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6961 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     7021 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     7081 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
