LOCUS Exported 6590 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3 flag p53 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6590) AUTHORS Gjoerup O, Zaveri D, Roberts TM TITLE Induction of p53-independent apoptosis by simian virus 40 small t antigen. JOURNAL J Virol. 2001 Oct . 75(19):9142-55. PUBMED 11533178 REFERENCE 2 (bases 1 to 6590) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6590 /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" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 907..930 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 937..2115 /codon_start=1 /gene="Homo sapiens TP53" /product="human tumor suppressor protein that is often mutated in cancers" /label=p53 /note="responds to cellular stresses to regulate the expression of target genes" /translation="EEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLS PDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQG SYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYK QSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPP EVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDR RTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMF RELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD" promoter complement(2143..2161) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2144..2161) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2181..2198) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2187..2411 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2457..2885 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2544..2563) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2754..2775 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2894..2914) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2899..3228 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3079..3214 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3141..3160 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3295..4089 /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(3349..3368) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3959..3978 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4263..4384 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4300..4319) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4354..4373 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4433..4449) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4433..4449) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4446..4468) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4457..4473 /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(4481..4511) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4526..4547 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4664..4681) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4835..5423) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4915..4934) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5594..6454) /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 6217..6236 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6455..6559) /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 ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc 901 gacatggact acaaggacga cgatgacaag ggatccgagg agccgcagtc agatcctagc 961 gtcgagcccc ctctgagtca ggaaacattt tcagacctat ggaaactact tcctgaaaac 1021 aacgttctgt cccccttgcc gtcccaagca atggatgatt tgatgctgtc cccggacgat 1081 attgaacaat ggttcactga agacccaggt ccagatgaag ctcccagaat gccagaggct 1141 gctccccccg tggcccctgc accagcagct cctacaccgg cggcccctgc accagccccc 1201 tcctggcccc tgtcatcttc tgtcccttcc cagaaaacct accagggcag ctacggtttc 1261 cgtctgggct tcttgcattc tgggacagcc aagtctgtga cttgcacgta ctcccctgcc 1321 ctcaacaaga tgttttgcca actggccaag acctgccctg tgcagctgtg ggttgattcc 1381 acacccccgc ccggcacccg cgtccgcgcc atggccatct acaagcagtc acagcacatg 1441 acggaggttg tgaggcgctg cccccaccat gagcgctgct cagatagcga tggtctggcc 1501 cctcctcagc atcttatccg agtggaagga aatttgcgtg tggagtattt ggatgacaga 1561 aacacttttc gacatagtgt ggtggtgccc tatgagccgc ctgaggttgg ctctgactgt 1621 accaccatcc actacaacta catgtgtaac agttcctgca tgggcggcat gaaccggagg 1681 cccatcctca ccatcatcac actggaagac tccagtggta atctactggg acggaacagc 1741 tttgaggtgc gtgtttgtgc ctgtcctggg agagaccggc gcacagagga agagaatctc 1801 cgcaagaaag gggagcctca ccacgagctg cccccaggga gcactaagcg agcactgccc 1861 aacaacacca gctcctctcc ccagccaaag aagaaaccac tggatggaga atatttcacc 1921 cttcagatcc gtgggcgtga gcgcttcgag atgttccgag agctgaatga ggccttggaa 1981 ctcaaggatg cccaggctgg gaaggagcca ggggggagca gggctcactc cagccacctg 2041 aagtccaaaa agggtcagtc tacctcccgc cataaaaaac tcatgttcaa gacagaaggg 2101 cctgactcag actgactcga gcatgcatct agagggccct attctatagt gtcacctaaa 2161 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 2221 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2281 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 2341 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 2401 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 2461 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 2521 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 2581 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 2641 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 2701 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 2761 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 2821 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 2881 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 3181 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 3241 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 3301 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3361 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 3421 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 3481 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 3541 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 3601 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3661 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3721 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3781 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 3841 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3901 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3961 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4021 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4081 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4141 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4201 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4261 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4321 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4381 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 4441 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4501 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4561 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 4621 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 4681 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 4741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 5041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 5101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 5161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 5221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 5281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 5341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 5401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 5461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 5521 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 5581 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 5641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 5701 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 5761 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 5821 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 5881 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 5941 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 6001 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 6061 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 6121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 6181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 6241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 6301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 6361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 6421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 6481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 6541 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc //