LOCUS Exported 6483 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Allows the integration of Myc ArA K162R in the genome and Tet-inducible expression. ACCESSION . VERSION . KEYWORDS pcDNA5 FRT TO Myc ArA K162R SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6483) AUTHORS Girdler F, Gascoigne KE, Eyers PA, Hartmuth S, Crafter C, Foote KM, Keen NJ, Taylor SS TITLE Validating Aurora B as an anti-cancer drug target. JOURNAL J Cell Sci. 2006 Sep 1;119(Pt 17):3664-75. Epub 2006 Aug 15. PUBMED 16912073 REFERENCE 2 (bases 1 to 6483) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6483 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 820..838 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 841..859 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 869..893 /label=LNCX /note="Human CMV promoter, forward primer" CDS 1154..1183 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2090..2101 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2435..2452) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2441..2665 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(2798..2817) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 2949..2996 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 3004..4026 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4156..4277 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4193..4212) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4247..4266 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4326..4342) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4326..4342) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4339..4361) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4350..4366 /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(4374..4404) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4419..4440 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4557..4574) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4728..5316) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4808..4827) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5487..6347) /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 6110..6129 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6348..6452) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttggtaccc cggggaattc ggcagatccc gaggtccgac 1021 agcgcccggc ccagatcccc acgcctgcca ggagcaagcc gagagccagc cggccggcgc 1081 actccgactc cgagcagtct ctgtccttcg acccgagccc cgcgcccttt ccgggacccc 1141 tgccccgcgg atggagcaga agctcatctc agaagaagac ctcgagggat ccgaccgatc 1201 taaagaaaac tgcatttcag gacctgttaa ggctacagct ccagttggag gtccaaaacg 1261 tgttctcgtg actcagcaat ttccttgtca gaatccatta cctgtaaata gtggccaggc 1321 tcagcgggtc ttgtgtcctt caaattcttc ccagcgcgtt cctttgcaag cacaaaagct 1381 tgtctccagt cacaagccgg ttcagaatca gaagcagaag caattgcagg caaccagtgt 1441 acctcatcct gtctccaggc cactgaataa cacccaaaag agcaagcagc ccctgccatc 1501 ggcacctgaa aataatcctg aggaggaact ggcatcaaaa cagaaaaatg aagaatcaaa 1561 aaagaggcag tgggctttgg aagactttga aattggtcgc cctctgggta aaggaaagtt 1621 tggtaatgtt tatttggcaa gagaaaagca aagcaagttt attctggctc ttagagtgtt 1681 atttaaagct cagctggaga aagccggagt ggagcatcag ctcagaagag aagtagaaat 1741 acagtcccac cttcggcatc ctaatattct tagactgtat ggttatttcc atgatgctac 1801 cagagtctac ctaattctgg aatatgcacc acttggaaca gtttatagag aacttcagaa 1861 actttcaaag tttgatgagc agagaactgc tacttatata acagaattgg caaatgccct 1921 gtcttactgt cattcgaaga gagttattca tagagacatt aagccagaga acttacttct 1981 tggatcagct ggagagctta aaattgcaga ttttgggtgg tcagtacatg ctccatcttc 2041 caggaggacc actctctgtg gcaccctgga ctacctgccc cctgaaatga ttgaaggtcg 2101 gatgcatgat gagaaggtgg atctctggag ccttggagtt ctttgctatg aatttttagt 2161 tgggaagcct ccttttgagg caaacacata ccaagagacc tacaaaagaa tatcacgggt 2221 tgaattcaca ttccctgact ttgtaacaga gggagccagg gacctcattt caagactgtt 2281 gaagcataat cccagccaga ggccaatgct cagagaagta cttgaacacc cctggatcac 2341 agcaaattca tcaaaaccat caaattgcca aaacaaagaa tcagctagca aacagtctta 2401 ggcggccgcg ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 2461 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2521 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 2581 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 2641 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 2701 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2761 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2821 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2881 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2941 cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg aacttccttg 3001 gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac 3061 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat 3121 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat 3181 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt 3241 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg 3301 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat 3361 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga 3421 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat 3481 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag 3541 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc 3601 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg 3661 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct 3721 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg 3781 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac 3841 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga 3901 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc 3961 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag 4021 gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4081 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4141 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4201 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4261 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4321 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4381 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4441 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4501 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4561 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 4621 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 4681 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 4741 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 4801 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 4861 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 4921 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 4981 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5041 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5101 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5161 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5221 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5281 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5341 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5401 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5461 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5521 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 5581 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 5641 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 5701 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 5761 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 5821 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 5881 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 5941 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6001 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6061 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6121 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6181 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6241 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6301 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6361 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6421 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6481 gtc //