LOCUS       sequence-240509-        8754 bp    DNA     circular SYN 14-APR-2025
DEFINITION  orange calcium-modulated bioluminescent indicator, 300nM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8754)
  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 8754)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_300
FEATURES             Location/Qualifiers
     source          1..8754
                     /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             3316..3393
                     /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             4258..4287
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4303..4320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4343..4360)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4349..4573
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4619..5047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4706..4725)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4916..4937
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5056..5076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5061..5390
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5241..5376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5303..5322
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5457..6251
                     /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(5511..5530)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6121..6140
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6427..6548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6464..6483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6518..6537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6597..6613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6597..6613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6610..6632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6621..6637
                     /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(6645..6651,6652..6669,6670..6675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6690..6711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6828..6845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6999..7587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7079..7098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7758..8549,8550..8618))
                     /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     8381..8400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8619..8723)
                     /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 gggggnnsgs gcgccrggcg gggsggggsg
      541 gggsgrgggg sggggsgggg sgrggcggag 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 agggtaagag gcgctggaag aaaaacttca ttgccgtcag cgctgccaac
     3361 cggttcaaga agatctccag ctccggggca ctggagctct ggaacggcaa caaaattatc
     3421 gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg
     3481 accggctggc ggctgtgcga acgcattctg gcgaggcacg aactcattaa agaaaatatg
     3541 cggtccaagc tctatctcga gggaagtgtg aatggtcatc aatttaagtg tactcatgag
     3601 ggcgaaggta aaccttacga aggaaagcaa acgaatcgaa taaaggtcgt ggaaggcgga
     3661 ccgctccctt ttgccttcga tattctggct acacatttca tgtatggatc taaagtcttt
     3721 attaagtatc ctgcggatct gccagactac tttaagcaga gctttccaga aggatttact
     3781 tgggaacgag tgatggtctt tgaggatgga ggtgtcctca cagctacgca agatacttcc
     3841 cttcaagatg gagaactgat ttacaacgtg aaagtgcgcg gtgtcaattt tcctgcaaat
     3901 ggtccagtca tgcaaaagaa gaccctcgga tgggaacctt ctacagaaac aatgtatcct
     3961 gcagatggag gtctcgaggg tcggtgtgat aaggctctta aactggtcgg aggaggtcat
     4021 ctccatgtga atttcaaaac tacctataaa agcaaaaagc ctgtcaaaat gccaggtgta
     4081 cattatgtcg atcgacggct cgagcggata aaggaagctg ataatgaaac gtatgtggaa
     4141 cagtatgaac atgccgtcgc acggtatagt aatctcggtg gcggaatgga tgaactctat
     4201 aagtaatgaa ttctgcagat atccagcaca gtggcggccg ctcgaggtca cccattcgaa
     4261 caaaaactca tctcagaaga ggatctgaat atgcataccg gtcatcatca ccatcaccat
     4321 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4381 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4441 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4501 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4561 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4621 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4681 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4741 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4801 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4861 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4921 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4981 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5041 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5161 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5221 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5281 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5341 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5401 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5461 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5521 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5581 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5641 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5701 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5761 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5821 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5881 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5941 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6001 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6061 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6121 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6181 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6241 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     6301 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6361 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6421 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6481 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6541 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     6601 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6661 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6721 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6781 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6841 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6901 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6961 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7021 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7081 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7141 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7201 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7261 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7321 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7381 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7441 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7501 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7561 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7621 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7681 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7741 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7801 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7861 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7921 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7981 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8041 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8101 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8161 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8221 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8281 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8341 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8401 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8461 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8521 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8581 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8641 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8701 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
