LOCUS Exported 5860 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of GFP-AURKA in mammalian cells. ACCESSION . VERSION . KEYWORDS pAuroraA-GFP-AURKA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5860) 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 5860) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5860 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 711..720 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 717..1433 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 717..1433 /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(762..783) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1023..1042) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1370..1391 /label=EGFP-C /note="EGFP, forward primer" CDS 2355..2366 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2796..2917 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2833..2852) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2887..2906 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2924..3379) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3061..3082) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3273..3292 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3406..3510 /gene="bla" /label=AmpR promoter promoter 3512..3869 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3535..3555) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3720..3855 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3782..3801 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3904..4698 /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(3958..3977) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4568..4587 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4886..4905) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4930..4977 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5306..34 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5795..5814 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 61 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 121 tgtggataac cgtattaccg ccatgcatta gttattaata gcttgacgtc aaggggctgt 181 tgcttcaccg ataaatggcc gaccgcggag agcaccctgg ggctgggact gccacaggtc 241 tggctggccg ttggctccac cacttccggg ttcttaggga gcaagtgcgc ctgcgcgcgg 301 tgtgcgccct taaacgcgac tcaaggcgtc gggtttgttg tcaaccaatc acaaggcagc 361 ctcgctcgag cgcaggccaa tcggctttct agctagaggg tttaactcct atttaaaaag 421 aagaaccttt gaattctaac ggctgagctc ttggaagact tgggtccttg ggtcgcaggt 481 gggagccgac gggtgggtag accgtggggg atatctcagt ggcggacgag gacggcgggg 541 acaaggggcg gctggtcgga gtggcggagc gtcaagtccc ctgtcggttc ctccgtccct 601 gagtgtcctt ggcgctgcct tgtgcccgcc cagcgccttt gcatccgctc ctgggcaccg 661 aggcgccctg taggatactg cttgttactt attacagcta gagaccggtc gccaccatgg 721 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 781 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 841 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 901 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 961 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 1021 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 1081 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 1141 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca 1201 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc 1261 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 1321 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc 1381 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac 1441 tcagatctcg agccatggac cgatctaaag aaaactgcat ttcaggacct gttaaggcta 1501 cagctccagt tggaggtcca aaacgtgttc tcgtgactca gcaatttcct tgtcagaatc 1561 cattacctgt aaatagtggc caggctcagc gggtcttgtg tccttcaaat tcttcccagc 1621 gcgttccttt gcaagcacaa aagcttgtct ccagtcacaa gccggttcag aatcagaagc 1681 agaagcaatt gcaggcaacc agtgtacctc atcctgtctc caggccactg aataacaccc 1741 aaaagagcaa gcagcccctg ccatcggcac ctgaaaataa tcctgaggag gaactggcat 1801 caaaacagaa aaatgaagaa tcaaaaaaga ggcagtgggc tttggaagac tttgaaattg 1861 gtcgccctct gggtaaagga aagtttggta atgtttattt ggcaagagaa aagcaaagca 1921 agtttattct ggctcttaaa gtgttattta aagctcagct ggagaaagcc ggagtggagc 1981 atcagctcag aagagaagta gaaatacagt cccaccttcg gcatcctaat attcttagac 2041 tgtatggtta tttccatgat gctaccagag tctacctaat tctggaatat gcaccacttg 2101 gaacagttta tagagaactt cagaaacttt caaagtttga tgagcagaga actgctactt 2161 atataacaga attggcaaat gcactatcat attgccattc gaagagagtt attcatagag 2221 acattaagcc agagaactta cttcttggat cagctggaga gcttaaaatt gcagattttg 2281 ggtggtcagt acatgctcca tcttccagga ggaccactct ctgtggcacc ctggactacc 2341 tgccccctga aatgattgaa ggtcggatgc atgatgagaa ggtggatctc tggagccttg 2401 gagttctttg ctatgaattt ttagttggga agcctccttt tgaggcaaac acataccaag 2461 agacctacaa aagaatatca cgggttgaat tcacattccc tgactttgta acagagggag 2521 ccagggacct catttcaaga ctgttgaagc ataatcccag ccagaggcca atgctcagag 2581 aagtacttga acacccctgg atcacagcaa attcatcaaa accatcaaat tgccaaaaca 2641 aagaatcagc tagcaaacag tcttagggat ccaccggatc tagataactg atcataatca 2701 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga 2761 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg 2821 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 2881 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat 2941 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 3001 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 3061 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 3121 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 3181 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 3241 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 3301 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 3361 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 3421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 3481 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg 3541 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3601 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 3661 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 3721 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 3781 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 3841 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt 3901 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3961 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4021 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 4081 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4141 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4201 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4261 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4321 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4381 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc 4441 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4501 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4561 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4621 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4681 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4741 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4801 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4861 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 4921 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata 4981 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg 5041 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc 5101 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata 5161 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 5221 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 5281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 5341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 5401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 5461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 5521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 5581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 5641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 5701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 5761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 5821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt //