LOCUS Exported 8142 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLenti6/V5-p53_R249S SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8142) AUTHORS Junk DJ, Vrba L, Watts GS, Oshiro MM, Martinez JD, Futscher BW TITLE Different mutant/wild-type p53 combinations cause a spectrum of increased invasive potential in nonmalignant immortalized human mammary epithelial cells. JOURNAL Neoplasia. 2008 May . 10(5):450-61. PUBMED 18472962 REFERENCE 2 (bases 1 to 8142) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8142 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 3..229 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 230..410 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 457..582 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1075..1308 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1493..1537 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" enhancer 1816..2119 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2120..2323 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2274..2294 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2320..2344 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2440..3618 /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="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML SPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP PEVGSDCTTIHYNYMCNSSCMGGMNRSPILTIITLEDSSGNLLGRNSFEVRVCACPGRD RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD" CDS 3652..3693 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3730..3750) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3735..4064 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3915..4050 /label=SV40 ori /note="SV40 origin of replication" promoter 4112..4159 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4178..4576 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" LTR 4663..4896 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4968..5089 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5005..5024) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5059..5078 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5129..5264 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5284..5303) /label=T7 /note="T7 promoter, forward primer" promoter complement(5285..5303) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5313..5330) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5313..5329) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5322..5344) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5471..5926 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5558..5577) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5768..5789 /label=F1ori-F /note="F1 origin, forward primer" promoter 5952..6056 /gene="bla" /label=AmpR promoter CDS 6057..6917 /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 complement(6275..6294) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7088..7676 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7577..7596 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7830..7847 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7964..7985 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8000..8030 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8038..8054 /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)." primer_bind 8043..8065 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8062..8078 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8062..8078 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8097..8117 /label=T3 /note="T3 promoter, forward primer" promoter 8099..8117 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca 61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga 121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt 181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg 241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg 421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt 481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg 541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga 601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta 661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta 721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga 781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg 841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt 901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga 961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc 1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc 1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct 1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag 1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca 1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg 1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa 1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgata 1801 agcttgggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 1861 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 1921 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 1981 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 2041 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 2101 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 2161 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 2221 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 2281 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca 2341 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gactctagag 2401 gatccactag tccagtgtgg tggaattgat cccttcacca tggaggagcc gcagtcagat 2461 cctagcgtcg agccccctct gagtcaggaa acattttcag acctatggaa actacttcct 2521 gaaaacaacg ttctgtcccc cttgccatcc caagcaatgg atgatttgat gctgtccccg 2581 gacgatattg aacaatggtt cactgaagac ccaggtccag atgaagctcc cagaatgcca 2641 gaggctgctc cccccgtggc ccctgcacca gcagctccta caccggcggc ccctgcacca 2701 gccccctcct ggcccctgtc atcttctgtc ccttcccaga aaacctacca gggcagctac 2761 ggtttccgtc tgggcttctt gcattctggg acagccaagt ctgtgacttg cacgtactcc 2821 cctgccctca acaagatgtt ttgccaactg gccaagacct gccctgtgca gctgtgggtt 2881 gattccacac ccccgcccgg cacccgcgtc cgcgccatgg ccatctacaa gcagtcacag 2941 cacatgacgg aggttgtgag gcgctgcccc caccatgagc gctgctcaga tagcgatggt 3001 ctggcccctc ctcagcatct tatccgagtg gaaggaaatt tgcgtgtgga gtatttggat 3061 gacagaaaca cttttcgaca tagtgtggtg gtgccctatg agccgcctga ggttggctct 3121 gactgtacca ccatccacta caactacatg tgtaacagtt cctgcatggg cggcatgaac 3181 cggagcccca tcctcaccat catcacactg gaagactcca gtggtaatct actgggacgg 3241 aacagctttg aggtgcgtgt ttgtgcctgt cctgggagag accggcgcac agaggaagag 3301 aatctccgca agaaagggga gcctcaccac gagctgcccc cagggagcac taagcgagca 3361 ctgcccaaca acaccagctc ctctccccag ccaaagaaga aaccactgga tggagaatat 3421 ttcacccttc agatccgtgg gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc 3481 ttggaactca aggatgccca ggctgggaag gagccagggg ggagcagggc tcactccagc 3541 cacctgaagt ccaaaaaggg tcagtctacc tcccgccata aaaaactcat gttcaagaca 3601 gaagggcctg actcagacaa gggctcgagt ctagagggcc cgcggttcga aggtaagcct 3661 atccctaacc ctctcctcgg tctcgattct acgcgtaccg gttagtaatg agtttggaat 3721 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 3781 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 3841 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 3901 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 3961 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc 4021 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt 4081 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat 4141 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga 4201 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga 4261 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt 4321 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc 4381 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag 4441 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt 4501 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta 4561 tgtgtgggag ggctaagcac aattcgagct cggtaccttt aagaccaatg acttacaagg 4621 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact 4681 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct 4741 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 4801 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 4861 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 4921 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg 4981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc 5101 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 5161 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 5221 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat 5281 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac 5341 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 5401 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 5461 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 5521 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 5581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 5641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 5701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 5761 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 5821 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 5881 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc 5941 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 6001 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga 6061 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 6121 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag 6181 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 6241 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 6301 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 6361 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 6421 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag 6481 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 6541 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 6601 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 6661 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 6721 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 6781 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 6841 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 6901 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 6961 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 7021 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 7081 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 7141 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 7201 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc 7261 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 7321 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 7381 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 7441 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 7501 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 7561 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 7621 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 7681 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 7741 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 7801 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 7861 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg 7921 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca 7981 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg 8041 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa 8101 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc tt //