LOCUS       sequence-320165-        6667 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 6667)
  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 6667)
  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 ShadowY-AURKA K162M-mTurq2
FEATURES             Location/Qualifiers
     source          1..6667
                     /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"
     misc_feature    1276..1341
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2257..2268
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(2587..2589,2590..2592,2593..3105)
                     /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    3395..3516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3432..3451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3486..3505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3523..3978)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3660..3681)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3872..3891
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4005..4109
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4111..4468
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4111..4302
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4134..4154)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4319..4454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4503..5297
                     /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             4503..5297
                     /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(4557..4576)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5485..5504)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5529..5576
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5905..6493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6394..6413
                     /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 gcttggcccg tggtgaatgt ctatatcatg
     1021 gccgacaagc agaagaacgg catcaaggtg aacttctcta tctatcataa catcgaggac
     1081 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cgggcccgtg
     1141 ctgctgcccg acaaccacta cctgagctac tcggctaagc tgagcaaaga ccccaacgag
     1201 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1261 gacgagctgt acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac
     1321 ggtaccgcgg gcccgggatc cggaagcttc gaattcatgg accgatctaa agaaaactgc
     1381 atttcaggac ctgttaaggc tacagctcca gttggaggtc caaaacgtgt tctcgtgact
     1441 cagcaatttc cttgtcagaa tccattacct gtaaatagtg gccaggctca gcgggtcttg
     1501 tgtccttcaa attcttccca gcgcattcct ttgcaagcac aaaaacttgt ctccagtcac
     1561 aagccggttc agaatcagaa gcagaagcaa ttgcaggcaa ccagtgtacc tcatcctgtc
     1621 tccaggccac tgaataacac ccaaaagagc aagcagcccc tgccatcggc acctgaaaat
     1681 aatcctgagg aggaactggc atcaaaacag aaaaatgaag aatcaaaaaa gaggcagtgg
     1741 gctttggaag actttgaaat tggtcgccct ctgggtaaag gaaagtttgg taatgtttat
     1801 ttggcaagag aaaagcaaag caagtttatt ctggctctta tggtgttatt taaagctcag
     1861 ctggagaaag ccggagtgga gcatcagctc agaagagaag tagaaataca gtcccacctt
     1921 cggcatccta atattcttag actgtatggt tatttccatg atgctaccag agtctaccta
     1981 attctggaat atgcaccact tggaacagtt tatagagaac ttcagaaact ttcaaagttt
     2041 gatgagcaga gaactgctac ttatataaca gaattggcaa atgcactatc atattgccat
     2101 tcgaagagag ttattcatag agacattaag ccagagaact tacttcttgg atcagctgga
     2161 gagcttaaaa ttgcagattt tgggtggtca gtacatgctc catcttccag gaggaccact
     2221 ctctgtggca ccctggacta cctgccccct gaaatgattg aaggtcggat gcatgatgag
     2281 aaggtggatc tctggagcct tggagttctt tgctatgaat ttttagttgg gaagcctcct
     2341 tttgaggcaa acacatacca agagacctac aaaagaatat cacgggttga gttcacattc
     2401 cctgactttg taacagaggg agccagggac ctcatttcaa gactgttgaa gcataatccc
     2461 agccagaggc caatgctcag agaagtactt gaacacccct ggatcacagc aaattcatca
     2521 aaaccatcaa attgccaaaa caaagaatca gctagcaaac agtctgggga tccgatggcg
     2581 gccgcaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2641 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2701 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2761 cccaccctcg tgaccaccct gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac
     2821 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2881 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2941 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     3001 gggcacaagc tggagtacaa ctactttagc gacaacgtct atatcaccgc cgacaagcag
     3061 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     3121 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     3181 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     3241 atggtcctgc tggagttcgt gaccgccgcc taagcggccg cgactctaga tcataatcag
     3301 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3361 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3421 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3481 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3541 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3601 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3661 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3721 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3781 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3841 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3901 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3961 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4021 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4081 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4141 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4201 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4261 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4321 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4381 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4441 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4501 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4561 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4621 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4681 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4741 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4801 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4861 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4921 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4981 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5041 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5101 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5161 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5221 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5281 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5341 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5401 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5461 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5521 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5581 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5641 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5701 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5761 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5821 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5881 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5941 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6001 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6061 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6121 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6181 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6241 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6301 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6361 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6421 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6481 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6541 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6601 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6661 agttccg
//
