LOCUS Exported 5894 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS GFP-p53 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5894) AUTHORS Boyd SD, Tsai KY, Jacks T TITLE An intact HDM2 RING-finger domain is required for nuclear exclusion of p53. JOURNAL Nat Cell Biol. 2000 Sep . 2(9):563-8. PUBMED 10980695 REFERENCE 2 (bases 1 to 5894) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5894 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 764..1067 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1068..1271 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1222..1242 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1343..2521 /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" regulatory 2537..2546 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2543..3262 /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 2543..3259 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2588..2609) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2849..2868) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3196..3217 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3385..3506 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3422..3441) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3476..3495 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3513..3968) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3650..3671) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3862..3881 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3995..4099 /gene="bla" /label=AmpR promoter promoter 4101..4458 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4124..4144) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4309..4444 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4371..4390 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4493..5287 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4547..4566) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5157..5176 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5475..5494) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5519..5566 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat 721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga 1321 gctcaagctt cgaattctgc agatggagga gccgcagtca gatcctagcg tcgagccccc 1381 tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc 1441 ccccttgccg tcccaagcaa tggatgattt gatgctgtcc ccggacgata ttgaacaatg 1501 gttcactgaa gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccgcgt 1561 ggcccctgca ccagcagctc ctacaccggc ggcccctgca ccagccccct cctggcccct 1621 gtcatcttct gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt 1681 cttgcattct gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat 1741 gttttgccaa ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc 1801 cggcacccgc gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt 1861 gaggcgctgc ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca 1921 tcttatccga gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg 1981 acatagtgtg gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca 2041 ctacaactac atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac 2101 catcatcaca ctggaagact ccagtggtaa tctactggga cggaacagct ttgaggtgcg 2161 tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg 2221 ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag 2281 ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg 2341 tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc 2401 ccaggctggg aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa 2461 gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc ctgactcaga 2521 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 2581 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 2641 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 2701 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 2761 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 2821 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 2881 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 2941 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 3001 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 3061 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 3121 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 3181 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 3241 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca 3301 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat 3361 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa 3421 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 3481 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa 3541 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 3601 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 3661 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 3721 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg 3781 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc 3841 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc 3901 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca 3961 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 4021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 4081 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta 4141 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 4201 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4261 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 4321 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 4381 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 4441 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga 4501 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 4561 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 4621 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga 4681 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 4741 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 4801 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 4861 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 4921 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 4981 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga 5041 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 5101 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 5161 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 5221 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 5281 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 5341 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 5401 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct 5461 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg 5521 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt 5581 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 5641 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 5701 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt 5761 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 5821 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 5881 atcaaaggat cttc //