LOCUS Exported 6270 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS hAur-A D274A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6270) AUTHORS Crane R, Kloepfer A, Ruderman JV TITLE Requirements for the destruction of human Aurora-A. JOURNAL J Cell Sci 2004 Dec 1;117(Pt 25):5975-83. Epub 2004 Nov 9. PUBMED 15536123 REFERENCE 2 (bases 1 to 6270) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6270 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 151..567 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 497..519 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 535..551 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1481..1492 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(1809..1829) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1814..2143 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1994..2129 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2056..2075 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2153..2752 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(2153..2172) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2649..2669 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 2973..3442 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(3572..3589) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3743..4331) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3823..3842) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4491..5351) /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 5114..5133 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5352..5456) /gene="bla" /label=AmpR promoter LTR 5487..5956 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 6021..6220 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" ORIGIN 1 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac 61 cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg 121 actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg 181 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag 241 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg 301 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct 361 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt 421 gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca 481 tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat 541 ccagccctca ctccttctct aggcgccggc cggatccacc atggaccgat ctaaagaaaa 601 ctgcatttca ggacctgtta aggctacagc tccagttgga ggtccaaaac gtgttctcgt 661 gactcagcaa tttccttgtc agaatccatt acctgtaaat agtggccagg ctcagcgggt 721 cttgtgtcct tcaaattctt cccagcgcgt tcctttgcaa gcacaaaagc ttgtctccag 781 tcacaagccg gttcagaatc agaagcagaa gcaattgcag gcaaccagtg tacctcatcc 841 tgtctccagg ccactgaata acacccaaaa gagcaagcag cccctgccat cggcacctga 901 aaataatcct gaggaggaac tggcatcaaa acagaaaaat gaagaatcaa aaaagaggca 961 gtgggctttg gaagactttg aaattggtcg ccctctgggt aaaggaaagt ttggtaatgt 1021 ttatttggca agagaaaagc aaagcaagtt tattctggct cttaaagtgt tatttaaagc 1081 tcagctggag aaagccggag tggagcatca gctcagaaga gaagtagaaa tacagtccca 1141 ccttcggcat cctaatattc ttagactgta tggttatttc catgatgcta ccagagtcta 1201 cctaattctg gaatatgcac cacttggaac agtttataga gaacttcaga aactttcaaa 1261 gtttgatgag cagagaactg ctacttatat aacagaattg gcaaatgccc tgtcttactg 1321 tcattcgaag agagttattc atagagacat taagccagag aacttacttc ttggatcagc 1381 tggagagctt aaaattgcag cttttgggtg gtcagtacat gctccatctt ccaggaggac 1441 cactctctgt ggcaccctgg actacctgcc ccctgaaatg attgaaggtc ggatgcatga 1501 tgagaaggtg gatctctgga gccttggagt tctttgctat gaatttttag ttgggaagcc 1561 tccttttgag gcaaacacat accaagagac ctacaaaaga atatcacggg ttgaattcac 1621 attccctgac tttgtaacag agggagccag ggacctcatt tcaagactgt tgaagcataa 1681 tcccagccag aggccaatgc tcagagaagt acttgaacac ccctggatca cagcaaattc 1741 atcaaaaccg tcaaattgcc aaaacaaaga atcagctagc aaacagtctt aggaatcctc 1801 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 1861 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 1921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 1981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 2041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca 2101 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga 2161 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 2221 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 2281 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 2341 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 2401 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 2461 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 2521 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 2581 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 2641 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 2701 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc 2761 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa 2821 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa 2881 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa 2941 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt 3001 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc 3061 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc 3121 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc 3181 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt 3241 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg 3301 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc 3361 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact 3421 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct 3481 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag 3541 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 3601 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 3661 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 3721 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 3781 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 3841 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 3901 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 3961 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4021 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4081 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4141 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 4201 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4261 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 4321 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 4381 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 4441 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 4501 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 4561 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 4621 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 4681 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 4741 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 4801 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 4861 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 4921 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 4981 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 5041 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 5101 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 5161 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 5221 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 5281 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 5341 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 5401 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 5461 tttccccgaa aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag 5521 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag 5581 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat 5641 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga 5701 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc 5761 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag 5821 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc 5881 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt 5941 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca 6001 ccgacccacc accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta 6061 gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc 6121 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt 6181 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat 6241 ccgaccccgt caggatatgt ggttctggta //