LOCUS       sequence-310867-        7662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AURKA fused to GFP at both ends in mammalian cells 
            under AurkA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7662)
  AUTHORS   Bertolin G, Sizaire F, Demeautis C, Chapuis C, Merola F, Erard M, 
            Tramier M
  TITLE     Optimized FRET Pairs and Quantification Approaches To Detect the 
            Activation of Aurora Kinase A at Mitosis.
  JOURNAL   ACS Sens. 2019 Aug 23;4(8):2018-2027. doi: 
            10.1021/acssensors.9b00486. Epub 2019 Jul 31.
  PUBMED    31317736
REFERENCE   2  (bases 1 to 7662)
  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"; doi: 
            "10.1021/acssensors.9b00486"; journalName: "ACS Sens"; date: 
            "2019-08-23- 23"; volume: "4"; issue: "8"; pages: "2018-2027"
COMMENT     Sequence Label: pAuroraA GFP-AURKA-GFP
FEATURES             Location/Qualifiers
     source          1..7662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             901..912
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(1255..1257,1258..1260,1261..1971)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1255..1257,1258..1260,1261..1971)
                     /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             1993..2022
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2038..2055
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2078..2095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2084..2308
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2354..2782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2441..2460)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2651..2672
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2791..2811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2796..3125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2976..3111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3038..3057
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3173..3220
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3239..3637
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3795..3916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3832..3851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3886..3905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3965..3981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3965..3981)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3978..4000)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3989..4005
                     /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(4013..4019,4020..4037,4038..4043))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4058..4079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4196..4213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4367..4955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4447..4466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5126..5917,5918..5986))
                     /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     5749..5768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5987..6091)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6166..6185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             join(6931..6933,6934..6936,6937..7647)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6931..6933,6934..6936,6937..7647)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 atggaccgat ctaaagaaaa ctgcatttca ggacctgtta aggctacagc tccagttgga
       61 ggtccaaaac gtgttctcgt gactcagcaa tttccttgtc agaatccatt acctgtaaat
      121 agtggccagg ctcagcgggt cttgtgtcct tcaaattctt cccagcgcat tcctttgcaa
      181 gcacaaaaac ttgtctccag tcacaagccg gttcagaatc agaagcagaa gcaattgcag
      241 gcaaccagtg tacctcatcc tgtctccagg ccactgaata acacccaaaa gagcaagcag
      301 cccctgccat cggcacctga aaataatcct gaggaggaac tggcatcaaa acagaaaaat
      361 gaagaatcaa aaaagaggca gtgggctttg gaagactttg aaattggtcg ccctctgggt
      421 aaaggaaagt ttggtaatgt ttatttggca agagaaaagc aaagcaagtt tattctggct
      481 cttaaagtgt tatttaaagc tcagctggag aaagccggag tggagcatca gctcagaaga
      541 gaagtagaaa tacagtccca ccttcggcat cctaatattc ttagactgta tggttatttc
      601 catgatgcta ccagagtcta cctaattctg gaatatgcac cacttggaac agtttataga
      661 gaacttcaga aactttcaaa gtttgatgag cagagaactg ctacttatat aacagaattg
      721 gcaaatgcac tatcatattg ccattcgaag agagttattc atagagacat taagccagag
      781 aacttacttc ttggatcagc tggagagctt aaaattgcag attttgggtg gtcagtacat
      841 gctccatctt ccaggaggac cactctctgt ggcaccctgg actacctgcc ccctgaaatg
      901 attgaaggtc ggatgcatga tgagaaggtg gatctctgga gccttggagt tctttgctat
      961 gaatttttag ttgggaagcc tccttttgag gcaaacacat accaagagac ctacaaaaga
     1021 atatcacggg ttgagttcac attccctgac tttgtaacag agggagccag ggacctcatt
     1081 tcaagactgt tgaagcataa tcccagccag aggccaatgc tcagagaagt acttgaacac
     1141 ccctggatca cagcaaattc atcaaaacca tcaaattgcc aaaacaaaga atcagctagc
     1201 aaacagtcca ctagtccagt gtggtggaat tctgcagata tccagcacag tggcatggtg
     1261 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1321 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1381 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1441 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1501 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1561 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1621 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1681 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1741 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1801 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1861 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1921 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gctcgagtct
     1981 agagggccct tcgaacaaaa actcatctca gaagaggatc tgaatatgca taccggtcat
     2041 catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2101 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2161 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2221 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2281 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2341 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2401 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2461 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2521 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2581 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2641 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2701 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2761 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2941 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3001 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3061 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3121 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     3181 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     3241 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     3301 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     3361 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     3421 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     3481 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     3541 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     3601 tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg
     3661 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3721 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3781 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3841 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3901 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3961 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4021 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4081 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4141 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4201 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4801 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5041 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5101 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5161 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5221 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5281 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5341 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5401 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5461 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5521 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5581 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5641 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5701 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5761 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5821 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5881 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5941 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6001 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6061 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     6181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     6241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     6301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata tcccctgatt
     6361 ctgtggataa ccgtattacc gccatgcatt agttattaat agcttgacgt caaggggctg
     6421 ttgcttcacc gataaatggc cgaccgcgga gagcaccctg gggctgggac tgccacaggt
     6481 ctggctggcc gttggctcca ccacttccgg gttcttaggg agcaagtgcg cctgcgcgcg
     6541 gtgtgcgccc ttaaacgcga ctcaaggcgt cgggtttgtt gtcaaccaat cacaaggcag
     6601 cctcgctcga gcgcaggcca atcggctttc tagctagagg gtttaactcc tatttaaaaa
     6661 gaagaacctt tgaattctaa cggctgagct cttggaagac ttgggtcctt gggtcgcagg
     6721 tgggagccga cgggtgggta gaccgtgggg gatatctcag tggcggacga ggacggcggg
     6781 gacaaggggc ggctggtcgg agtggcggag cgtcaagtcc cctgtcggtt cctccgtccc
     6841 tgagtgtcct tggcgctgcc ttgtgcccgc ccagcgcctt tgcatccgct cctgggcacc
     6901 ggtaccgagc tcggattggt accgagctcg atggtgagca agggcgagga gctgttcacc
     6961 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     7021 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     7081 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     7141 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     7201 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     7261 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     7321 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     7381 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     7441 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     7501 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     7561 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     7621 actctcggca tggacgagct gtacaaggaa ttcgagctca cc
//
