LOCUS Exported 6649 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mammalian expression of human p53 S15A mutant. . ACCESSION . VERSION . KEYWORDS pcDNA3 p53 S15A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6649) AUTHORS Loughery J, Cox M, Smith LM, Meek DW TITLE Critical role for p53-serine 15 phosphorylation in stimulating transactivation at p53-responsive promoters. JOURNAL Nucleic Acids Res. 2014 Jul;42(12):7666-80. doi: 10.1093/nar/gku501. Epub 2014 Jun 13. PUBMED 24928858 REFERENCE 2 (bases 1 to 6649) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6649 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(2204..2222) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2205..2222) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2242..2259) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2248..2472 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2518..2946 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2605..2624) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2815..2836 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2955..2975) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2960..3289 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3140..3275 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3202..3221 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3356..4150 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3410..3429) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4020..4039 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4324..4445 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4361..4380) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4415..4434 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4494..4510) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4494..4510) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4507..4529) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4518..4534 /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(4542..4572) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4587..4608 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4725..4742) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4896..5484) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4976..4995) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5655..6515) /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 6278..6297 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6516..6620) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta 901 ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattcggctt atggaggagc 961 cgcagtcaga tcctagcgtc gagcccccgc tagctcagga aacattttca gacctatgga 1021 aactacttcc tgaaaacaac gttctgtccc ccttgccgtc ccaagcaatg gatgatttga 1081 tgctgtcccc ggacgatatt gaacaatggt tcactgaaga cccaggtcca gatgaagctc 1141 ccagaatgcc agaggctgct ccccccgtgg cccctgcacc agcagctcct acaccggcgg 1201 cccctgcacc agccccctcc tggcccctgt catcttctgt cccttcccag aaaacctacc 1261 agggcagcta cggtttccgt ctgggcttct tgcattctgg gacagccaag tctgtgactt 1321 gcacgtactc ccctgccctc aacaagatgt tttgccaact ggccaagacc tgccctgtgc 1381 agctgtgggt tgattccaca cccccgcccg gcacccgcgt ccgcgccatg gccatctaca 1441 agcagtcaca gcacatgacg gaggttgtga ggcgctgccc ccaccatgag cgctgctcag 1501 atagcgatgg tctggcccct cctcagcatc ttatccgagt ggaaggaaat ttgcgtgtgg 1561 agtatttgga tgacagaaac acttttcgac atagtgtggt ggtgccctat gagccgcctg 1621 aggttggctc tgactgtacc accatccact acaactacat gtgtaacagt tcctgcatgg 1681 gcggcatgaa ccggaggccc atcctcacca tcatcacact ggaagactcc agtggtaatc 1741 tactgggacg gaacagcttt gaggtgcgtg tttgtgcctg tcctgggaga gaccggcgca 1801 cagaggaaga gaatctccgc aagaaagggg agcctcacca cgagctgccc ccagggagca 1861 ctaagcgagc actgcccaac aacaccagct cctctcccca gccaaagaag aaaccactgg 1921 atggagaata tttcaccctt cagatccgtg ggcgtgagcg cttcgagatg ttccgagagc 1981 tgaatgaggc cttggaactc aaggatgccc aggctgggaa ggagccaggg gggagcaggg 2041 ctcactccag ccacctgaag tccaaaaagg gtcagtctac ctcccgccat aaaaaactca 2101 tgttcaagac agaagggcct gactcagact gatctagaaa gccgaattct gcagatatcc 2161 atcacactcc cgggcgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa 2221 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 2281 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 2341 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 2401 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 2461 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 2521 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 2581 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 2641 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 2701 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 2761 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 2821 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 2881 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 2941 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 3001 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 3061 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 3121 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 3181 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 3241 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 3301 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat 3361 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta 3421 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca 3481 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga 3541 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga 3601 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct 3661 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg 3721 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga 3781 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca 3841 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga 3901 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg 3961 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc 4021 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt 4081 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga 4141 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca 4201 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc 4261 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac 4321 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 4381 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 4441 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag 4501 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 4561 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 4621 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 4681 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 4741 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4801 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4861 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4921 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4981 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5041 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 5101 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 5161 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5221 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5281 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 5341 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5401 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5461 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5521 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5581 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5761 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 5821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5941 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 6001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 6061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 6121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6241 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6481 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6601 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg //