LOCUS       sequence-240525-        8763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  orange calcium-modulated bioluminescent indicator, 110nM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8763)
  AUTHORS   Oh Y, Park Y, Cho JH, Wu H, Paulk NK, Liu LX, Kim N, Kay MA, Wu JC, 
            Lin MZ
  TITLE     An orange calcium-modulated bioluminescent indicator for 
            non-invasive activity imaging.
  JOURNAL   Nat Chem Biol. 2019 Apr 1. pii: 10.1038/s41589-019-0256-z. doi: 
            10.1038/s41589-019-0256-z.
  PUBMED    30936501
REFERENCE   2  (bases 1 to 8763)
  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 Chem 
            Biol. 2019 Apr 1. pii: 10.1038/s41589-019-0256-z. doi: 
            10.1038/s41589-019-0256-z."
COMMENT     Sequence Label: pcDNA3.1-Orange_CaMBI_110
FEATURES             Location/Qualifiers
     source          1..8763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        92..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          674..1690
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1614..1636
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1698..1717
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1780..1789
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3325..3402
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             4267..4296
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4312..4329
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4352..4369)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4358..4582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4628..5056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4715..4734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4925..4946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5065..5085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5070..5399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5250..5385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5312..5331
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5466..6260
                     /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(5520..5539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6130..6149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6436..6557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6473..6492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6527..6546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6606..6622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6606..6622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6619..6641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6630..6646
                     /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(6654..6660,6661..6678,6679..6684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6699..6720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6837..6854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7008..7596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7088..7107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7767..8558,8559..8627))
                     /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     8390..8409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8628..8732)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcaatt ctgcaggaat ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      421 tcactctccc catctccccc cccctcccca cccccaattt tgtatttatt tattttttaa
      481 ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgccrggsg gggsggggsg
      541 gggsgrgggg sggggsgggg cgrggcggag aggtgcggcg gcagccaatc agagcggcgc
      601 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      661 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      721 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      781 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      841 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
      901 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
      961 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
     1021 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
     1081 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     1141 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     1201 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     1261 gccgggcggg gcggggccgc ctcgggccgg ggrgggctcg ggggaggggc gcggcggccc
     1321 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     1381 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     1441 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     1501 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     1561 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1621 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     1681 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1741 attgatttga taccgcgggc ccgggatcag cttggtaccg ccaccatggt gagcaagggc
     1801 gaggagctga tcaaggagaa catgagaagc aagctgtacc tggaaggcag cgtgaacggc
     1861 caccagttca agtgcaccca cgaaggggag ggcaagccct acgagggcaa gcagaccaac
     1921 aggatcaagg tggtggaggg aggccccctg ccgttcgcat tcgacatcct ggccacccac
     1981 tttatgtacg ggagcaaggt gttcatcaag taccccgccg acctccccga ttattttaag
     2041 cagtccttcc ctgagggctt cacatgggag agagtcatgg tgttcgaaga cgggggcgtg
     2101 ctgaccgcca cccaggacac cagcctccag gacggcgagc tcatctacaa cgtcaaggtc
     2161 agaggggtga acttcccagc caacggcccc gtgatgcaga agaaaacact gggctgggag
     2221 cccagcaccg agaccatgta ccccgctgac ggcggcctgg aaggcagatg cgacaaggcc
     2281 ctgaagctcg tgggcggggg ccacctgcac gtcaacttca agaccacata caagtccaag
     2341 aaacccgtga agatgcccgg cgtccactac gtggaccgca gactggaaag aatcaaggag
     2401 gccgacaacg agacctacgt cgagcagtac gagcacgctg tggccagata ctccaacctg
     2461 ggcggaggct tcacactcga agatttcgtt ggggactggc gacagacagc cggctacaac
     2521 ctggaccaag tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc
     2581 gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat
     2641 gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt
     2701 aaggtggtgt accctgtgga tgatcatcac tttaaggtga tcctgcacta tggcacactg
     2761 gtaatcgacg gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc
     2821 gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgattatgca tgaccaactg
     2881 acagaagagc agattgcaga gttcaaagaa gccttctcat tattcgacaa ggatggggac
     2941 ggcaccatca ccacaaagga acttggcacc gttatgaggt cgcttggaca aaacccaacg
     3001 gaagcagaat tgcaggatat gatcaatgaa gtcgatgctg atggcaatgg aacgatttac
     3061 tttcctgaat ttcttactat gatggctaga aaaatgaagg acacagacag cgaagaggaa
     3121 atccgagaag cattccgtgt ttttgacaag gatgggaacg gctacatcag cgctgctcag
     3181 ttacgtcacg tcatgacaaa cctcggggag aagttaacag atgaagaagt tgatgaaatg
     3241 ataagggaag cagatatcga tggtgatggc caagtaaact atgaagagtt tgtacaaatg
     3301 atgacagcaa agggtggcgg gtcaaagagg cgctggaaga aaaacttcat tgccgtcagc
     3361 gctgccaacc ggttcaagaa gatctccagc tccggggcac tggagctctg gaacggcaac
     3421 aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc
     3481 aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgaggcacga actcattaaa
     3541 gaaaatatgc ggtccaagct ctatctcgag ggaagtgtga atggtcatca atttaagtgt
     3601 actcatgagg gcgaaggtaa accttacgaa ggaaagcaaa cgaatcgaat aaaggtcgtg
     3661 gaaggcggac cgctcccttt tgccttcgat attctggcta cacatttcat gtatggatct
     3721 aaagtcttta ttaagtatcc tgcggatctg ccagactact ttaagcagag ctttccagaa
     3781 ggatttactt gggaacgagt gatggtcttt gaggatggag gtgtcctcac agctacgcaa
     3841 gatacttccc ttcaagatgg agaactgatt tacaacgtga aagtgcgcgg tgtcaatttt
     3901 cctgcaaatg gtccagtcat gcaaaagaag accctcggat gggaaccttc tacagaaaca
     3961 atgtatcctg cagatggagg tctcgagggt cggtgtgata aggctcttaa actggtcgga
     4021 ggaggtcatc tccatgtgaa tttcaaaact acctataaaa gcaaaaagcc tgtcaaaatg
     4081 ccaggtgtac attatgtcga tcgacggctc gagcggataa aggaagctga taatgaaacg
     4141 tatgtggaac agtatgaaca tgccgtcgca cggtatagta atctcggtgg cggaatggat
     4201 gaactctata agtaatgaat tctgcagata tccagcacag tggcggccgc tcgaggtcac
     4261 ccattcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg tcatcatcac
     4321 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4381 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4441 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4501 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4561 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4621 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4681 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4741 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4801 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4861 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4921 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4981 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5041 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5101 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5221 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5281 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5341 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5401 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5461 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5521 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5581 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5641 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5701 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5761 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5821 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5881 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5941 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6001 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6061 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6121 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6181 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6241 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6301 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6361 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6421 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6481 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6541 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6601 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6661 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6721 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6781 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6841 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6901 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6961 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7021 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7201 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7441 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7621 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7681 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7741 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7801 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7861 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7921 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7981 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8041 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8101 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8161 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8221 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8281 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8341 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8401 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8461 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8521 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8581 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8641 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8701 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8761 gtc
//
