LOCUS Exported 6550 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of GFP-AURKA K162M-mCherry in cells. ACCESSION . VERSION . KEYWORDS pAuroraA-GFP-AURKA_K162M-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6550) AUTHORS Bertolin G, Sizaire F, Herbomel G, Reboutier D, Prigent C, Tramier M TITLE A FRET biosensor reveals spatiotemporal activation and functions of aurora kinase A in living cells. JOURNAL Nat Commun. 2016 Sep 14;7:12674. doi: 10.1038/ncomms12674. PUBMED 27624869 REFERENCE 2 (bases 1 to 6550) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6550 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 624..1340 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 624..1340 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(669..690) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(930..949) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1277..1298 /label=EGFP-C /note="EGFP, forward primer" CDS 2256..2267 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" CDS 2577..3284 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(2722..2740) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2990..3009 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 3432..3553 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3469..3488) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3523..3542 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3560..4015) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3697..3718) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3909..3928 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4042..4146 /gene="bla" /label=AmpR promoter promoter 4148..4505 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4171..4191) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4356..4491 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4418..4437 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4540..5334 /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(4594..4613) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5204..5223 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5522..5541) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5566..5613 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5942..6530 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6431..6450 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 61 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagctt gacgtcaagg 121 ggctgttgct tcaccgataa atggccgacc gcggagagca ccctggggct gggactgcca 181 caggtctggc tggccgttgg ctccaccact tccgggttct tagggagcaa gtgcgcctgc 241 gcgcggtgtg cgcccttaaa cgcgactcaa ggcgtcgggt ttgttgtcaa ccaatcacaa 301 ggcagcctcg ctcgagcgca ggccaatcgg ctttctagct agagggttta actcctattt 361 aaaaagaaga acctttgaat tctaacggct gagctcttgg aagacttggg tccttgggtc 421 gcaggtggga gccgacgggt gggtagaccg tgggggatat ctcagtggcg gacgaggacg 481 gcggggacaa ggggcggctg gtcggagtgg cggagcgtca agtcccctgt cggttcctcc 541 gtccctgagt gtccttggcg ctgccttgtg cccgcccagc gcctttgcat ccgctcctgg 601 gcaccggtac cgagctcgga tccatggtga gcaagggcga ggagctgttc accggggtgg 661 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 721 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 781 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca 841 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 901 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 961 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1021 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1081 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 1141 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 1201 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca 1261 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 1321 gcatggacga gctgtacaag gaattcgagc tcaccatgga ccgatctaaa gaaaactgca 1381 tttcaggacc tgttaaggct acagctccag ttggaggtcc aaaacgtgtt ctcgtgactc 1441 agcaatttcc ttgtcagaat ccattacctg taaatagtgg ccaggctcag cgggtcttgt 1501 gtccttcaaa ttcttcccag cgcattcctt tgcaagcaca aaaacttgtc tccagtcaca 1561 agccggttca gaatcagaag cagaagcaat tgcaggcaac cagtgtacct catcctgtct 1621 ccaggccact gaataacacc caaaagagca agcagcccct gccatcggca cctgaaaata 1681 atcctgagga ggaactggca tcaaaacaga aaaatgaaga atcaaaaaag aggcagtggg 1741 ctttggaaga ctttgaaatt ggtcgccctc tgggtaaagg aaagtttggt aatgtttatt 1801 tggcaagaga aaagcaaagc aagtttattc tggctcttat ggtgttattt aaagctcagc 1861 tggagaaagc cggagtggag catcagctca gaagagaagt agaaatacag tcccaccttc 1921 ggcatcctaa tattcttaga ctgtatggtt atttccatga tgctaccaga gtctacctaa 1981 ttctggaata tgcaccactt ggaacagttt atagagaact tcagaaactt tcaaagtttg 2041 atgagcagag aactgctact tatataacag aattggcaaa tgcactatca tattgccatt 2101 cgaagagagt tattcataga gacattaagc cagagaactt acttcttgga tcagctggag 2161 agcttaaaat tgcagatttt gggtggtcag tacatgctcc atcttccagg aggaccactc 2221 tctgtggcac cctggactac ctgccccctg aaatgattga aggtcggatg catgatgaga 2281 aggtggatct ctggagcctt ggagttcttt gctatgaatt tttagttggg aagcctcctt 2341 ttgaggcaaa cacataccaa gagacctaca aaagaatatc acgggttgag ttcacattcc 2401 ctgactttgt aacagaggga gccagggacc tcatttcaag actgttgaag cataatccca 2461 gccagaggcc aatgctcaga gaagtacttg aacacccctg gatcacagca aattcatcaa 2521 aaccatcaaa ttgccaaaac aaagaatcag ctagcaaaca gtctgtcgac aagcttatgg 2581 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc 2641 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 2701 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg 2761 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg 2821 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga 2881 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg 2941 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc 3001 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc 3061 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg 3121 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca 3181 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg 3241 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagctcgag tctagaggat 3301 aaggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt 3361 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 3421 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 3481 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 3541 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat 3601 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa 3661 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta 3721 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca 3781 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat 3841 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg 3901 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc 3961 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt 4021 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 4081 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 4141 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt 4201 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 4261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4321 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 4381 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 4441 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 4501 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 4561 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 4621 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 4681 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta 4741 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 4801 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 4861 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 4921 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 4981 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 5041 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc 5101 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 5161 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 5221 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 5281 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 5341 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 5401 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 5461 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa 5521 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac 5581 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct 5641 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct 5701 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg 5761 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc 5821 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 5881 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 5941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 6001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 6061 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 6121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 6181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 6241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 6301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 6361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 6421 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 6481 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 6541 acgcggcctt //