LOCUS       sequence-309886-        6542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of kinase-dead AURKA K62M  in mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6542)
  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 6542)
  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: pcDNA 3.1  mitoAURKA Lys162Met
FEATURES             Location/Qualifiers
     source          1..6542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             683..757
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1520..1531
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1899..1916)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1905..2129
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2175..2603
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2262..2281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2472..2493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2612..2632)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2617..2946
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2797..2932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2859..2878
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3013..3807
                     /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(3067..3086)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     polyA_signal    3981..4102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4018..4037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4072..4091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4151..4167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4151..4167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4164..4186)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4175..4191
                     /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(4199..4205,4206..4223,4224..4229))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4244..4265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4382..4399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4553..5141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4633..4652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5312..6103,6104..6172))
                     /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     5935..5954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6173..6277)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6352..6371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttatgtccgt cctgacgccg ctgctgctgc ggggcttgac
      721 aggctcggcc cggcggctcc cagtgccgcg cgccaagatc cattcgttgt caaattcttc
      781 ccagcgcgtt cctttgcaag cacaaaagct tgtctccagt cacaagccgg ttcagaatca
      841 gaagcagaag caattgcagg caaccagtgt acctcatcct gtctccaggc cactgaataa
      901 cacccaaaag agcaagcagc ccctgccatc ggcacctgaa aataatcctg aggaggaact
      961 ggcatcaaaa cagaaaaatg aagaatcaaa aaagaggcag tgggctttgg aagactttga
     1021 aattggtcgc cctctgggta aaggaaagtt tggtaatgtt tatttggcaa gagaaaagca
     1081 aagcaagttt attctggctc ttatggtgtt atttaaagct cagctggaga aagccggagt
     1141 ggagcatcag ctcagaagag aagtagaaat acagtcccac cttcggcatc ctaatattct
     1201 tagactgtat ggttatttcc atgatgctac cagagtctac ctaattctgg aatatgcacc
     1261 acttggaaca gtttatagag aacttcagaa actttcaaag tttgatgagc agagaactgc
     1321 tacttatata acagaattgg caaatgcact atcatattgc cattcgaaga gagttattca
     1381 tagagacatt aagccagaga acttacttct tggatcagct ggagagctta aaattgcaga
     1441 ttttgggtgg tcagtacatg ctccatcttc caggaggacc actctctgtg gcaccctgga
     1501 ctacctgccc cctgaaatga ttgaaggtcg gatgcatgat gagaaggtgg atctctggag
     1561 ccttggagtt ctttgctatg aatttttagt tgggaagcct ccttttgagg caaacacata
     1621 ccaagagacc tacaaaagaa tatcacgggt tgaattcaca ttccctgact ttgtaacaga
     1681 gggagccagg gacctcattt caagactgtt gaagcataat cccagccaga ggccaatgct
     1741 cagagaagta cttgaacacc cctggatcac agcaaattca tcaaaaccat caaattgcca
     1801 aaacaaagaa tcagctagca aacagtctga attctgcaga tatccagcac agtggcggcc
     1861 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     1921 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1981 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2041 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2101 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2161 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2221 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2281 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2341 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2401 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2461 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2521 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2581 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     2641 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2701 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2761 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2821 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2881 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2941 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3001 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3061 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3121 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3181 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3241 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3301 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3361 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3421 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3481 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3541 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3601 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3661 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3721 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3781 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     3841 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3901 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     3961 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4021 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4081 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4141 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4201 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4261 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4321 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4381 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4441 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4501 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4561 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4621 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4681 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4741 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4801 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4861 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4921 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4981 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5041 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5101 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5161 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5221 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5281 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5341 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5401 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5461 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5521 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5581 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5641 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5701 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5761 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5821 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5881 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5941 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6001 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6061 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6121 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6181 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6241 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6301 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     6361 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     6421 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     6481 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     6541 tt
//
