LOCUS Exported 6869 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBI-p53 wt/EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6869) AUTHORS Yu J, Zhang L, Hwang PM, Rago C, Kinzler KW, Vogelstein B TITLE Identification and classification of p53-regulated genes. JOURNAL Proc Natl Acad Sci U S A. 1999 Dec 7. 96(25):14517-22. PUBMED 10588737 REFERENCE 2 (bases 1 to 6869) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6869 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 370..389 /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" polyA_signal 388..443 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind 489..508 /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" intron 644..1216 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" CDS complement(1785..2966) /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 SPDDIEQWFTEDPGPDEAPRMPEAAPRVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD" primer_bind complement(3044..3060) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3045..3061) /label=pBluescriptKS /note="For pBluescript vector" promoter complement(3201..3239) /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 3272..3290 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3314..3332 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3356..3374 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3398..3416 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 3440..3458 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 3601..3617 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 3601..3617 /label=pBluescriptSK /note="For pBluescript vector" CDS 3625..4344 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3625..4341 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3670..3691) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3931..3950) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4278..4299 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4381..4515 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4405..4424) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4459..4478 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(4878..4896) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4964..5068 /gene="bla" /label=AmpR promoter CDS 5069..5929 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5287..5306) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6100..6688 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6589..6608 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6842..6859 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagtcgag 61 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa 121 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa 181 acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata aggaatggac 241 agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt tcatggcagc 301 cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat gttttaaatg 361 atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac acactattgc 421 aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct tcatgatgtc 481 cccataattt ttggcagagg gaaaaagatc gatctcagtg gtatttgtga gccagggcat 541 tggccacacc agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattctttgc 601 caaaatgatg agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga 661 aggcatgaac atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct 721 caaacagtgt atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt 781 tgtttctacc aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac 841 cttatcattt aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac 901 cctgattgcc ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa 961 aagtgaattt aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa 1021 atgaaaagaa aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa 1081 attatcatga gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga 1141 aaactttgcc ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa 1201 gggtccccaa actcaccctg aagttctcag ctctagagat atcgtcgagg ccgcgctagc 1261 gtttaaactg tcgaccttaa ttaagagatc ccgctagacc ccatgtaata aaaggtggtt 1321 tcaaggccag atgtacatta tttcattaac cctcacaatg cactctgtga ggtaggtgca 1381 aatgccagca tttcacagat atgggccttg aagttagaga aaattcaaca gtgagggaca 1441 gcttccctgg ttagtacggt gaagtgggcc cctacctaga atgtggctga ttgtaaacta 1501 acccttaact gcaagaacat ttcttacatc tcccaaacat ccctcacagt aaaaacctta 1561 aaatctaagc tggtatgtcc tactccccat cctcctcccc acaacaaaac accagtgcag 1621 gccaacttgt tcagtggagc cccgggacaa agcaaatgga agtcctgggt gcttctgacg 1681 cacacctatt gcaagcaagg gttcaaagac ccaaaaccca aaatggcagg ggagggagag 1741 atgggggtgg gaggctgtca gtggggaaca agaagtggag aatgtcagtc tgagtcaggc 1801 ccttctgtct tgaacatgag ttttttatgg cgggaggtag actgaccctt tttggacttc 1861 aggtggctgg agtgagccct gctcccccct ggctccttcc cagcctgggc atccttgagt 1921 tccaaggcct cattcagctc tcggaacatc tcgaagcgct cacgcccacg gatctgaagg 1981 gtgaaatatt ctccatccag tggtttcttc tttggctggg gagaggagct ggtgttgttg 2041 ggcagtgctc gcttagtgct ccctgggggc agctcgtggt gaggctcccc tttcttgcgg 2101 agattctctt cctctgtgcg ccggtctctc ccaggacagg cacaaacacg cacctcaaag 2161 ctgttccgtc ccagtagatt accactggag tcttccagtg tgatgatggt gaggatgggc 2221 ctccggttca tgccgcccat gcaggaactg ttacacatgt agttgtagtg gatggtggta 2281 cagtcagagc caacctcagg cggctcatag ggcaccacca cactatgtcg aaaagtgttt 2341 ctgtcatcca aatactccac acgcaaattt ccttccactc ggataagatg ctgaggaggg 2401 gccagaccat cgctatctga gcagcgctca tggtgggggc agcgcctcac aacctccgtc 2461 atgtgctgtg actgcttgta gatggccatg gcgcggacgc gggtgccggg cgggggtgtg 2521 gaatcaaccc acagctgcac agggcaggtc ttggccagtt ggcaaaacat cttgttgagg 2581 gcaggggagt acgtgcaagt cacagacttg gctgtcccag aatgcaagaa gcccagacgg 2641 aaaccgtagc tgccctggta ggttttctgg gaagggacag aagatgacag gggccaggag 2701 ggggctggtg caggggccgc cggtgtagga gctgctggtg caggggccac gcggggagca 2761 gcctctggca ttctgggagc ttcatctgga cctgggtctt cagtgaacca ttgttcaata 2821 tcgtccgggg acagcatcaa atcatccatt gcttgggacg gcaaggggga cagaacgttg 2881 ttttcaggaa gtagtttcca taggtctgaa aatgtttcct gactcagagg gggctcgacg 2941 ctaggatctg actgcggctc ctccatggca gtgacccgga aggcagtctg gctgccaatc 3001 cagggaagcg tgtcaccgtc gtggaattcg atatcaagct tatcgatacc gtcgacctcg 3061 aggcgcgtca gctgactaga ggatccccgg gtaccgagct cgaattcggg gccgcggagg 3121 ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg tctccaggcg 3181 atctgacggt tcactaaacg agctctgctt atataggcct cccaccgtac acgcctacct 3241 cgacccgggt accgagctcg actttcactt ttctctatca ctgataggga gtggtaaact 3301 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct 3361 atcactgata gggagtggta aactcgactt tcacttttct ctatcactga tagggagtgg 3421 taaactcgac tttcactttt ctctatcact gatagggagt ggtaaactcg acctatataa 3481 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 3541 tccatagaag acaccgggac cgatccagcc tccgcggccc cgaattcctg cagcggccgc 3601 tctagaacta gtggatcccc gcggatggtg agcaagggcg aggagctgtt caccggggtg 3661 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 3721 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 3781 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc 3841 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 3901 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 3961 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 4021 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat 4081 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc 4141 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 4201 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc 4261 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc 4321 ggcatggacg agctgtacaa gtaagaattc gatatcaagc ttgtcgacct agttctagag 4381 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga 4441 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc 4501 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag 4561 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat 4621 cctgcaagcc tcgtcgtcct ggccggacca cgctatctgt gcaaggtccc cggccccgga 4681 cgcgcgctcc atgagcagag cgcccgccgc cgaggcgaag actcgggcgg cgccctgccc 4741 gtcccaccag gtcaacaggc ggtaaccggc ctcttcatcg ggaatgcgcg cgaccttcag 4801 catcgccggc atgtccccct ggcggacggg aagtatccag ctcgaccaag ctggccgcgt 4861 cgagtgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 4921 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 4981 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 5041 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 5101 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 5161 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 5221 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 5281 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 5341 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 5401 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 5461 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 5521 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 5581 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 5641 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 5701 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 5761 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 5821 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 5881 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 5941 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 6001 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 6061 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 6121 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 6181 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 6241 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 6301 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 6361 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 6421 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 6481 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 6541 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 6601 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 6661 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 6721 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 6781 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 6841 cagcgagtca gtgagcgagg aagcggaag //