LOCUS       sequence-310878-        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"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(559..561,562..564,565..1275)
                     /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             2191..2202
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(2521..2523,2524..2526,2527..3039)
                     /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    3329..3450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3366..3385)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3420..3439
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3457..3912)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3594..3615)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3806..3825
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3939..4043
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4045..4402
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4045..4236
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4068..4088)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4253..4388
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4437..5231
                     /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             4437..5231
                     /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(4491..4510)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5419..5438)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5463..5510
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5839..6427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6328..6347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      601 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      661 gagggcgatg ccacctacgg caagctgacc ctgaagctca tctgcaccac cggcaagctg
      721 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcgcgc
      781 tatccagacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      841 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      901 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      961 ggcaacatcc tggggcacaa gctcgagtac aactggaaca gccacaacgt ctatatcatg
     1021 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1081 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1141 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     1201 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1261 gacgagctgt acaagggaag cttcgaattc atggaccgat ctaaagaaaa ctgcatttca
     1321 ggacctgtta aggctacagc tccagttgga ggtccaaaac gtgttctcgt gactcagcaa
     1381 tttccttgtc agaatccatt acctgtaaat agtggccagg ctcagcgggt cttgtgtcct
     1441 tcaaattctt cccagcgcat tcctttgcaa gcacaaaaac ttgtctccag tcacaagccg
     1501 gttcagaatc agaagcagaa gcaattgcag gcaaccagtg tacctcatcc tgtctccagg
     1561 ccactgaata acacccaaaa gagcaagcag cccctgccat cggcacctga aaataatcct
     1621 gaggaggaac tggcatcaaa acagaaaaat gaagaatcaa aaaagaggca gtgggctttg
     1681 gaagactttg aaattggtcg ccctctgggt aaaggaaagt ttggtaatgt ttatttggca
     1741 agagaaaagc aaagcaagtt tattctggct cttatggtgt tatttaaagc tcagctggag
     1801 aaagccggag tggagcatca gctcagaaga gaagtagaaa tacagtccca ccttcggcat
     1861 cctaatattc ttagactgta tggttatttc catgatgcta ccagagtcta cctaattctg
     1921 gaatatgcac cacttggaac agtttataga gaacttcaga aactttcaaa gtttgatgag
     1981 cagagaactg ctacttatat aacagaattg gcaaatgcac tatcatattg ccattcgaag
     2041 agagttattc atagagacat taagccagag aacttacttc ttggatcagc tggagagctt
     2101 aaaattgcag attttgggtg gtcagtacat gctccatctt ccaggaggac cactctctgt
     2161 ggcaccctgg actacctgcc ccctgaaatg attgaaggtc ggatgcatga tgagaaggtg
     2221 gatctctgga gccttggagt tctttgctat gaatttttag ttgggaagcc tccttttgag
     2281 gcaaacacat accaagagac ctacaaaaga atatcacggg ttgagttcac attccctgac
     2341 tttgtaacag agggagccag ggacctcatt tcaagactgt tgaagcataa tcccagccag
     2401 aggccaatgc tcagagaagt acttgaacac ccctggatca cagcaaattc atcaaaacca
     2461 tcaaattgcc aaaacaaaga atcagctagc aaacagtctg gggatccgat ggcggccgca
     2521 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2581 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     2641 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2701 ctcgtgacca ccctgtcctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     2761 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2821 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2881 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2941 aagctggagt acaactactt tagcgacaac gtctatatca ccgccgacaa gcagaagaac
     3001 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
     3061 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3121 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3181 ctgctggagt tcgtgaccgc cgcctaagcg gccgcgactc tagatcataa tcagccatac
     3241 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3301 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3361 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3421 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3481 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3541 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3601 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3661 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3721 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3781 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3841 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3901 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3961 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4021 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4081 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4141 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4201 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4261 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4321 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4381 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4441 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4501 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4561 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4621 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4681 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4741 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4801 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4861 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4921 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4981 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5041 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5101 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5161 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5221 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5281 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5341 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5401 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5461 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5521 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5581 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5641 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5701 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5761 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6481 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6541 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6601 g
//
