LOCUS       sequence-309889-        5756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AURKA without Nter, fused to GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5756)
  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 5756)
  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: pEGFP N1 AURKA deltaNter
FEATURES             Location/Qualifiers
     source          1..5756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             189..983
                     /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             189..983
                     /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(243..262)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(1171..1190)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1215..1262
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1591..2179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2080..2099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2354..2657
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2658..2861
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2812..2832
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3665..3676
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3989..3998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3995..3997,3998..4000,4001..4714)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3995..3997,3998..4000,4001..4711)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4040..4061)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4301..4320)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4648..4669
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4837..4958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4874..4893)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4928..4947
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4965..5420)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5102..5123)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5314..5333
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5447..5551
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5553..5756,1..154)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5553..5744
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5576..5596)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
       61 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      121 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
      181 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      241 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
      301 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
      361 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
      421 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
      481 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
      541 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
      601 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
      661 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
      721 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
      781 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
      841 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
      901 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
      961 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     1021 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     1081 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     1141 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     1201 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     1261 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     1321 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     1381 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     1441 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1501 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1561 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1621 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1681 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1741 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1801 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1861 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1921 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1981 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2041 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2101 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2161 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2221 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2281 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     2341 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2401 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2461 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2521 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2581 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2641 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2701 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2761 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2821 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     2881 tccgctagcg ctaccggact cagatctcga gatgtcaaat tcttcccagc gcgttccttt
     2941 gcaagcacaa aagcttgtct ccagtcacaa gccggttcag aatcagaagc agaagcaatt
     3001 gcaggcaacc agtgtacctc atcctgtctc caggccactg aataacaccc aaaagagcaa
     3061 gcagcccctg ccatcggcac ctgaaaataa tcctgaggag gaactggcat caaaacagaa
     3121 aaatgaagaa tcaaaaaaga ggcagtgggc tttggaagac tttgaaattg gtcgccctct
     3181 gggtaaagga aagtttggta atgtttattt ggcaagagaa aagcaaagca agtttattct
     3241 ggctcttaaa gtgttattta aagctcagct ggagaaagcc ggagtggagc atcagctcag
     3301 aagagaagta gaaatacagt cccaccttcg gcatcctaat attcttagac tgtatggtta
     3361 tttccatgat gctaccagag tctacctaat tctggaatat gcaccacttg gaacagttta
     3421 tagagaactt cagaaacttt caaagtttga tgagcagaga actgctactt atataacaga
     3481 attggcaaat gcactatcat attgccattc gaagagagtt attcatagag acattaagcc
     3541 agagaactta cttcttggat cagctggaga gcttaaaatt gcagattttg ggtggtcagt
     3601 acatgctcca tcttccagga ggaccactct ctgtggcacc ctggactacc tgccccctga
     3661 aatgattgaa ggtcggatgc atgatgagaa ggtggatctc tggagccttg gagttctttg
     3721 ctatgaattt ttagttggga agcctccttt tgaggcaaac acataccaag agacctacaa
     3781 aagaatatca cgggttgaat tcacattccc tgactttgta acagagggag ccagggacct
     3841 catttcaaga ctgttgaagc ataatcccag ccagaggcca atgctcagag aagtacttga
     3901 acacccctgg atcacagcaa attcatcaaa accatcaaat tgccaaaaca aagaatcagc
     3961 tagcaaacag tctggggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     4021 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4081 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4141 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     4201 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4261 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4321 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4381 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4441 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4501 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4561 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4621 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4681 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc
     4741 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4801 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4861 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4921 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     4981 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5041 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5101 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5161 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5221 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5281 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5341 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5401 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5461 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5521 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     5581 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5641 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5701 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactcc
//
