LOCUS       sequence-304559-        6601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AuroraA kinase-dead biosensor under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6601)
  AUTHORS   Bertolin G, Bulteau AL, Alves-Guerra MC, Burel A, Lavault MT, Gavard
            O, Le Bras S, Gagne JP, Poirier GG, Le Borgne R, Prigent C, Tramier 
            M
  TITLE     Aurora kinase A localises to mitochondria to control organelle 
            dynamics and energy production.
  JOURNAL   eLife. 2018 Aug 2;7. pii: 38111. doi: 10.7554/eLife.38111.
  PUBMED    30070631
REFERENCE   2  (bases 1 to 6601)
  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: "eLife."; 
            date: "2018-08-2"; volume: "7. pii: 38111. doi"; pages: " 
            10.7554/eLife.38111"
COMMENT     Sequence Label: pCMV ShadowG-AURKA K162M-mTurq2
FEATURES             Location/Qualifiers
     source          1..6601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      36..171
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             220..1014
                     /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             220..1014
                     /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(274..293)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(1202..1221)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1246..1293
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1622..2210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2111..2130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2385..2688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2689..2892
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2843..2863
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2943..2945,2946..2948,2949..3659)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4575..4586
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(4905..4907,4908..4910,4911..5423)
                     /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"
     polyA_signal    5713..5834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5750..5769)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5804..5823
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5841..6296)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5978..5999)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6190..6209
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6323..6427
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6429..6601,1..185)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(6429..6601,1..19)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6452..6472)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
       61 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      121 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      181 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
      241 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
      301 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
      361 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
      421 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
      481 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
      541 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
      601 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
      661 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
      721 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
      781 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
      841 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
      901 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
      961 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     1021 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     1081 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     1141 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     1201 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     1261 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     1321 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     1381 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     1441 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     1501 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1561 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1621 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1681 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1741 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     1801 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1861 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1921 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1981 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2041 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2101 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2161 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2221 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2281 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     2341 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     2401 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     2461 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     2521 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     2581 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     2641 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     2701 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     2761 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2821 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2881 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg
     2941 agatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3001 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3061 acggcaagct gaccctgaag ctcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3121 ccctcgtgac caccttcggc tacggcctga tgtgcttcgc gcgctatcca gaccacatga
     3181 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3241 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3301 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3361 acaagctcga gtacaactgg aacagccaca acgtctatat catggccgac aagcagaaga
     3421 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3481 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3541 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     3601 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     3661 gaagcttcga attcatggac cgatctaaag aaaactgcat ttcaggacct gttaaggcta
     3721 cagctccagt tggaggtcca aaacgtgttc tcgtgactca gcaatttcct tgtcagaatc
     3781 cattacctgt aaatagtggc caggctcagc gggtcttgtg tccttcaaat tcttcccagc
     3841 gcattccttt gcaagcacaa aaacttgtct ccagtcacaa gccggttcag aatcagaagc
     3901 agaagcaatt gcaggcaacc agtgtacctc atcctgtctc caggccactg aataacaccc
     3961 aaaagagcaa gcagcccctg ccatcggcac ctgaaaataa tcctgaggag gaactggcat
     4021 caaaacagaa aaatgaagaa tcaaaaaaga ggcagtgggc tttggaagac tttgaaattg
     4081 gtcgccctct gggtaaagga aagtttggta atgtttattt ggcaagagaa aagcaaagca
     4141 agtttattct ggctcttaaa gtgttattta aagctcagct ggagaaagcc ggagtggagc
     4201 atcagctcag aagagaagta gaaatacagt cccaccttcg gcatcctaat attcttagac
     4261 tgtatggtta tttccatgat gctaccagag tctacctaat tctggaatat gcaccacttg
     4321 gaacagttta tagagaactt cagaaacttt caaagtttga tgagcagaga actgctactt
     4381 atataacaga attggcaaat gcactatcat attgccattc gaagagagtt attcatagag
     4441 acattaagcc agagaactta cttcttggat cagctggaga gcttaaaatt gcagattttg
     4501 ggtggtcagt acatgctcca tcttccagga ggaccactct ctgtggcacc ctggactacc
     4561 tgccccctga aatgattgaa ggtcggatgc atgatgagaa ggtggatctc tggagccttg
     4621 gagttctttg ctatgaattt ttagttggga agcctccttt tgaggcaaac acataccaag
     4681 agacctacaa aagaatatca cgggttgagt tcacattccc tgactttgta acagagggag
     4741 ccagggacct catttcaaga ctgttgaagc ataatcccag ccagaggcca atgctcagag
     4801 aagtacttga acacccctgg atcacagcaa attcatcaaa accatcaaat tgccaaaaca
     4861 aagaatcagc tagcaaacag tctggggatc cgatggcggc cgcaatggtg agcaagggcg
     4921 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4981 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     5041 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgt
     5101 cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     5161 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     5221 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     5281 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     5341 actttagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     5401 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     5461 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     5521 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     5581 ccgccgccta agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     5641 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     5701 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5761 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5821 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     5881 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5941 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6001 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6061 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6121 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6181 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6241 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     6301 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6361 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6421 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     6481 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6541 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6601 t
//
