LOCUS Exported 5906 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS p53-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5906) TITLE p53-GFP REFERENCE 2 (bases 1 to 5906) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5906 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 16..319 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 320..523 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 474..494 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 601..610 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 607..1785 /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 1801..1810 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1807..2526 /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 1807..2523 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1852..1873) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2113..2132) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2460..2481 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2649..2770 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2686..2705) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2740..2759 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2777..3232) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2914..2935) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3126..3145 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3259..3363 /gene="bla" /label=AmpR promoter promoter 3365..3722 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3388..3408) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3573..3708 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3635..3654 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3757..4551 /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(3811..3830) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4421..4440 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4739..4758) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4783..4830 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5159..5747 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5648..5667 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca 541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacc 601 gccaccatgg aggagccgca gtcagatcct agcgtcgagc cccctctgag tcaggaaaca 661 ttttcagacc tatggaaact acttcctgaa aacaacgttc tgtccccctt gccgtcccaa 721 gcaatggatg atttgatgct gtccccggac gatattgaac aatggttcac tgaagaccca 781 ggtccagatg aagctcccag aatgccagag gctgctcccc gcgtggcccc tgcaccagca 841 gctcctacac cggcggcccc tgcaccagcc ccctcctggc ccctgtcatc ttctgtccct 901 tcccagaaaa cctaccaggg cagctacggt ttccgtctgg gcttcttgca ttctgggaca 961 gccaagtctg tgacttgcac gtactcccct gccctcaaca agatgttttg ccaactggcc 1021 aagacctgcc ctgtgcagct gtgggttgat tccacacccc cgcccggcac ccgcgtccgc 1081 gccatggcca tctacaagca gtcacagcac atgacggagg ttgtgaggcg ctgcccccac 1141 catgagcgct gctcagatag cgatggtctg gcccctcctc agcatcttat ccgagtggaa 1201 ggaaatttgc gtgtggagta tttggatgac agaaacactt ttcgacatag tgtggtggtg 1261 ccctatgagc cgcctgaggt tggctctgac tgtaccacca tccactacaa ctacatgtgt 1321 aacagttcct gcatgggcgg catgaaccgg aggcccatcc tcaccatcat cacactggaa 1381 gactccagtg gtaatctact gggacggaac agctttgagg tgcgtgtttg tgcctgtcct 1441 gggagagacc ggcgcacaga ggaagagaat ctccgcaaga aaggggagcc tcaccacgag 1501 ctgcccccag ggagcactaa gcgagcactg cccaacaaca ccagctcctc tccccagcca 1561 aagaagaaac cactggatgg agaatatttc acccttcaga tccgtgggcg tgagcgcttc 1621 gagatgttcc gagagctgaa tgaggccttg gaactcaagg atgcccaggc tgggaaggag 1681 ccagggggga gcagggctca ctccagccac ctgaagtcca aaaagggtca gtctacctcc 1741 cgccataaaa aactcatgtt caagacagaa gggcctgact cagacgggga tccaccggtc 1801 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag 1861 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc 1921 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg 1981 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac 2041 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc 2101 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac 2161 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg 2221 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag 2281 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag 2341 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac 2401 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac 2461 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac 2521 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac 2581 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg 2641 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 2701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 2761 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt 2821 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca 2881 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta 2941 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta 3001 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg 3061 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga 3121 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg 3181 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc 3241 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 3301 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 3361 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3541 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3721 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 3781 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 3841 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 3901 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg 3961 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4021 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4081 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 4141 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4201 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 4261 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat 4321 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 4381 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 4441 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 4501 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 4561 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 4621 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 4681 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg 4741 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga 4801 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc 4861 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt 4921 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg 4981 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt 5041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 5101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 5161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 5221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 5281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 5341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 5401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 5461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 5521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 5581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 5641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 5701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 5761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 5821 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca 5881 attacggggt cattagttca tagccc //