LOCUS Exported 6550 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of GFP-AURKA-mCherry in mammalian cells. ACCESSION . VERSION . KEYWORDS pAuroraA-GFP-AURKA-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 650..1366 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 650..1366 /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(695..716) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(956..975) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1303..1324 /label=EGFP-C /note="EGFP, forward primer" CDS 2282..2293 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" CDS 2603..3310 /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(2748..2766) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3016..3035 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 3458..3579 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3495..3514) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3549..3568 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3586..4041) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3723..3744) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3935..3954 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4068..4172 /gene="bla" /label=AmpR promoter promoter 4174..4531 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4197..4217) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4382..4517 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4444..4463 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4566..5360 /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(4620..4639) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5230..5249 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5548..5567) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5592..5639 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5968..6 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6457..6476 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 61 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat 121 tagttattaa tagcttgacg tcaaggggct gttgcttcac cgataaatgg ccgaccgcgg 181 agagcaccct ggggctggga ctgccacagg tctggctggc cgttggctcc accacttccg 241 ggttcttagg gagcaagtgc gcctgcgcgc ggtgtgcgcc cttaaacgcg actcaaggcg 301 tcgggtttgt tgtcaaccaa tcacaaggca gcctcgctcg agcgcaggcc aatcggcttt 361 ctagctagag ggtttaactc ctatttaaaa agaagaacct ttgaattcta acggctgagc 421 tcttggaaga cttgggtcct tgggtcgcag gtgggagccg acgggtgggt agaccgtggg 481 ggatatctca gtggcggacg aggacggcgg ggacaagggg cggctggtcg gagtggcgga 541 gcgtcaagtc ccctgtcggt tcctccgtcc ctgagtgtcc ttggcgctgc cttgtgcccg 601 cccagcgcct ttgcatccgc tcctgggcac cggtaccgag ctcggatcca tggtgagcaa 661 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 721 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac 781 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 841 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt 901 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga 961 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 1021 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 1081 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt 1141 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca 1201 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac 1261 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt 1321 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaaggaat tcgagctcac 1381 catggaccga tctaaagaaa actgcatttc aggacctgtt aaggctacag ctccagttgg 1441 aggtccaaaa cgtgttctcg tgactcagca atttccttgt cagaatccat tacctgtaaa 1501 tagtggccag gctcagcggg tcttgtgtcc ttcaaattct tcccagcgca ttcctttgca 1561 agcacaaaaa cttgtctcca gtcacaagcc ggttcagaat cagaagcaga agcaattgca 1621 ggcaaccagt gtacctcatc ctgtctccag gccactgaat aacacccaaa agagcaagca 1681 gcccctgcca tcggcacctg aaaataatcc tgaggaggaa ctggcatcaa aacagaaaaa 1741 tgaagaatca aaaaagaggc agtgggcttt ggaagacttt gaaattggtc gccctctggg 1801 taaaggaaag tttggtaatg tttatttggc aagagaaaag caaagcaagt ttattctggc 1861 tcttaaagtg ttatttaaag ctcagctgga gaaagccgga gtggagcatc agctcagaag 1921 agaagtagaa atacagtccc accttcggca tcctaatatt cttagactgt atggttattt 1981 ccatgatgct accagagtct acctaattct ggaatatgca ccacttggaa cagtttatag 2041 agaacttcag aaactttcaa agtttgatga gcagagaact gctacttata taacagaatt 2101 ggcaaatgca ctatcatatt gccattcgaa gagagttatt catagagaca ttaagccaga 2161 gaacttactt cttggatcag ctggagagct taaaattgca gattttgggt ggtcagtaca 2221 tgctccatct tccaggagga ccactctctg tggcaccctg gactacctgc cccctgaaat 2281 gattgaaggt cggatgcatg atgagaaggt ggatctctgg agccttggag ttctttgcta 2341 tgaattttta gttgggaagc ctccttttga ggcaaacaca taccaagaga cctacaaaag 2401 aatatcacgg gttgagttca cattccctga ctttgtaaca gagggagcca gggacctcat 2461 ttcaagactg ttgaagcata atcccagcca gaggccaatg ctcagagaag tacttgaaca 2521 cccctggatc acagcaaatt catcaaaacc atcaaattgc caaaacaaag aatcagctag 2581 caaacagtct gtcgacaagc ttatggtgag caagggcgag gaggataaca tggccatcat 2641 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga 2701 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt 2761 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg 2821 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc 2881 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac 2941 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa 3001 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga 3061 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa 3121 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca 3181 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta 3241 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga 3301 gctgtacaag ctcgagtcta gaggataagg atccaccgga tctagataac tgatcataat 3361 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct 3421 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa 3481 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 3541 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta 3601 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg 3661 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg 3721 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa 3781 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg 3841 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt 3901 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg 3961 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta 4021 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 4081 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 4141 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa 4201 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 4261 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 4321 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 4381 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 4441 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 4501 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt 4561 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 4621 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 4681 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 4741 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 4801 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 4861 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 4921 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 4981 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 5041 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc 5101 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 5161 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 5221 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 5281 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 5341 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc 5401 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 5461 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 5521 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac 5581 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca 5641 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg 5701 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg 5761 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata 5821 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 5881 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 5941 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 6001 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 6061 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 6121 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 6181 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 6241 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 6301 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 6361 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 6421 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 6481 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 6541 gcggagccta //