LOCUS       sequence-176637-        6778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan color variant of bright luminescent protein, targeted to the 
            Mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6778)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 6778)
  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 
            Commun."; date: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: Cox8x2-CeNL_pcDNA3
FEATURES             Location/Qualifiers
     source          1..6778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     766..785
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        766..784
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             818..892
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             914..985
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(1007..1009,1010..1012,1013..1525)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(1052..1073)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1313..1332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1660..1681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2234..2252)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2235..2252)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2272..2289)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2278..2502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2548..2976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2635..2654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2845..2866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2985..3005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2990..3319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3170..3305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3232..3251
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3386..4180
                     /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(3440..3459)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4050..4069
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4354..4475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4391..4410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4445..4464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4524..4540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4524..4540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4537..4559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4548..4564
                     /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(4572..4578,4579..4596,4597..4602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4617..4638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4755..4772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4926..5514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5006..5025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5685..6476,6477..6545))
                     /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     6308..6327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6546..6650)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6725..6744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
       61 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
      121 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      661 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta
      781 tagggagacc caagcttgaa ccgtcagatc cgctagcatg tccgtcctga cgccgctgct
      841 gctgcggggc ttgacaggct cggcccggcg gctcccagtg ccgcgcgcca agatccattc
      901 gttgggggat ctgtccgtcc tgacgccgct gctgctgcgg ggcttgacag gctcggcccg
      961 gcggctccca gtgccgcgcg ccaagatcca ttcgttgggg gatccgatgg tgagcaaggg
     1021 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1081 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1141 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1201 gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1261 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1321 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1381 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1441 ctactttagc gacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     1501 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     1561 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1621 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1681 gaccgccgcc gggttgcata cactcgaaga tttcgttggg gactggcgac agacagccgg
     1741 ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg
     1801 ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga
     1861 catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa
     1921 aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg
     1981 cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga
     2041 aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa
     2101 caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat
     2161 caacggagtg accggctggc ggctgtgcga acgcattctg gcgtaactcg agcatgcatc
     2221 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2281 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2341 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2401 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2461 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2521 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2761 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2881 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2941 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3001 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3061 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3121 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3181 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3241 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3301 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3361 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3421 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3481 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3541 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3601 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3661 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3721 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3781 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3841 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3901 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3961 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4021 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4081 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4141 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4201 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4261 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4321 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4501 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4561 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4621 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4681 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4741 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4801 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4861 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4981 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5041 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5101 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5161 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5221 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5281 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5341 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5401 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5461 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5521 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5581 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5641 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5701 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5761 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5821 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5881 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5941 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6001 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6061 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6121 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6181 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6241 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6301 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6361 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6421 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6481 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6541 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6661 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     6721 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgt
//
