LOCUS       sequence-139142-        8318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8318)
  AUTHORS   Baker SJ, Markowitz S, Fearon ER, Willson JK, Vogelstein B
  TITLE     Suppression of human colorectal carcinoma cell growth by wild-type 
            p53.
  JOURNAL   Science. 1990 Aug 24. 249(4971):912-5.
  PUBMED    2144057
REFERENCE   2  (bases 1 to 8318)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            1990 Aug 24. 249(4971):912-5."
COMMENT     Sequence Label: pCMV-Neo-Bam p53 R249S
FEATURES             Location/Qualifiers
     source          1..8318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     561..580
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    579..634
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     680..699
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1288..2469)
                     /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"
     intron          2671..3243
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3334..3358)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(3355..3558)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3384..3404)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3559..3938
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4231..4278
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4303..4322
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4516..5310)
                     /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(4627..4646)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5237..5256
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5367..5512)
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(5923..5940)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5923..5939)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5932..5954)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6148..6167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6267..6289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6327..6345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6413..6517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6518..6586,6587..7378)
                     /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(6736..6755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7549..8137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8038..8057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8291..8308
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattgcggcc
      241 gatccgtcga gggatctcca taagagaaga gggacagcta tgactgggag tagtcaggag
      301 aggaggaaaa atctggctag taaaacatgt aaggaaaatt ttagggatgt taaagaaaaa
      361 aataacacaa aacaaaatat aaaaaaaatc taacctcaag tcaaggcttt tctatggaat
      421 aaggaatgga cagcaggggg ctgtttcata tactgatgac ctctttatag ccaacctttg
      481 ttcatggcag ccagcatatg ggcatatgtt gccaaactct aaaccaaata ctcattctga
      541 tgttttaaat gatttgccct cccatatgtc cttccgagtg agagacacaa aaaattccaa
      601 cacactattg caatgaaaat aaatttcctt tattagccag aagtcagatg ctcaaggggc
      661 ttcatgatgt ccccataatt tttggcagag ggaaaaagat ctcagtggta tttgtgagcc
      721 agggcattgg ccacaccagc caccaccttc tgataggcag cctgcacctg aggagtgaat
      781 tcctcgacgg atcccgctag accccatgta ataaaaggtg gtttcaaggc cagatgtaca
      841 ttatttcatt aaccctcaca atgcactctg tgaggtaggt gcaaatgcca gcatttcaca
      901 gatatgggcc ttgaagttag agaaaattca acagtgaggg acagcttccc tggttagtac
      961 ggtgaagtgg gcccctacct agaatgtggc tgattgtaaa ctaaccctta actgcaagaa
     1021 catttcttac atctcccaaa catccctcac agtaaaaacc ttaaaatcta agctggtatg
     1081 tcctactccc catcctcctc cccacaacaa aacaccagtg caggccaact tgttcagtgg
     1141 agccccggga caaagcaaat ggaagtcctg ggtgcttctg acgcacacct attgcaagca
     1201 agggttcaaa gacccaaaac ccaaaatggc aggggaggga gagatggggg tgggaggctg
     1261 tcagtgggga acaagaagtg gagaatgtca gtctgagtca ggcccttctg tcttgaacat
     1321 gagtttttta tggcgggagg tagactgacc ctttttggac ttcaggtggc tggagtgagc
     1381 cctgctcccc cctggctcct tcccagcctg ggcatccttg agttccaagg cctcattcag
     1441 ctctcggaac atctcgaagc gctcacgccc acggatctga agggtgaaat attctccatc
     1501 cagtggtttc ttctttggct ggggagagga gctggtgttg ttgggcagtg ctcgcttagt
     1561 gctccctggg ggcagctcgt ggtgaggctc ccctttcttg cggagattct cttcctctgt
     1621 gcgccggtct ctcccaggac aggcacaaac acgcacctca aagctgttcc gtcccagtag
     1681 attaccactg gagtcttcca gtgtgatgat ggtgaggatg ggactccggt tcatgccgcc
     1741 catgcaggaa ctgttacaca tgtagttgta gtggatggtg gtacagtcag agccaacctc
     1801 aggcggctca tagggcacca ccacactatg tcgaaaagtg tttctgtcat ccaaatactc
     1861 cacacgcaaa tttccttcca ctcggataag atgctgagga ggggccagac catcgctatc
     1921 tgagcagcgc tcatggtggg ggcagcgcct cacaacctcc gtcatgtgct gtgactgctt
     1981 gtagatggcc atggcgcgga cgcgggtgcc gggcgggggt gtggaatcaa cccacagctg
     2041 cacagggcag gtcttggcca gttggcaaaa catcttgttg agggcagggg agtacgtgca
     2101 agtcacagac ttggctgtcc cagaatgcaa gaagcccaga cggaaaccgt agctgccctg
     2161 gtaggttttc tgggaaggga cagaagatga caggggccag gagggggctg gtgcaggggc
     2221 cgccggtgta ggagctgctg gtgcaggggc cacgggggga gcagcctctg gcattctggg
     2281 agcttcatct ggacctgggt cttcagtgaa ccattgttca atatcgtccg gggacagcat
     2341 caaatcatcc attgcttggg acggcaaggg ggacagaacg ttgttttcag gaagtagttt
     2401 ccataggtct gaaaatgttt cctgactcag agggggctcg acgctaggat ctgactgcgg
     2461 ctcctccatg gcagtgaccc ggaaggcagt ctggctgcca atccagggaa gcgtgtcacc
     2521 gtcgtggaaa gcacgctccc agcccgaacg caaagtgtcc ccggagccca gcagctacct
     2581 gctccctgga cggtggctct agcgggatcc gtcgaggaat tctttgccaa aatgatgaga
     2641 cagcacaaca accagcacgt tgcccaggag ctgtaggaaa gagaagaagg catgaacatg
     2701 gttagcagag gggcccggtt tggactcaga gtattttatc ctcatctcaa acagtgtata
     2761 tcattgtaac cataaagaga aaggcaggat gatgaccagg gtgtagttgt ttctaccaat
     2821 aagaatattt ccacgccagc cagaatttat atgcagaaat attctacctt atcatttaat
     2881 tataacaatt gttctctaaa actgtgctga agtacaatat aatataccct gattgccttg
     2941 aaaaaaaagt gattagagaa agtacttaca atctgacaaa taaacaaaag tgaatttaaa
     3001 aattcgttac aaatgcaagc taaagtttaa cgaaaaagtt acagaaaatg aaaagaaaat
     3061 aagaggagac aatggttgtc aacagagtag aaagtgaaag aaacaaaatt atcatgaggg
     3121 tccatggtga tacaagggac atcttcccat tctaaacaac accctgaaaa ctttgccccc
     3181 tccatataac atgaatttta caatagcgaa aaagaaagaa caatcaaggg tccccaaact
     3241 caccctgaag ttctcaggat cggtcgaccg gaggctggat cggtcccggt gtcttctatg
     3301 gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct
     3361 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     3421 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     3481 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     3541 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     3601 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     3661 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     3721 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     3781 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     3841 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     3901 aattgattac tattaataac tagtcaataa tcaatgtcaa catggcggta atgttggaca
     3961 tgagccaata taaatgtaca tattatgata tggatacaac gtatgcaatg ggccaagctc
     4021 tctagagggc cgcgatccgc cgtccctaac ccacggggcc cgtggctatg gcagggcttg
     4081 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg
     4141 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt
     4201 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat
     4261 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt
     4321 cagttagcct cccccatctc ccggggtggg cgaagaactc cagcatgaga tccccgcgct
     4381 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     4441 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
     4501 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     4561 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     4621 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     4681 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     4741 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
     4801 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     4861 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     4921 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     4981 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     5041 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     5101 cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt
     5161 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     5221 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     5281 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     5341 agatctgcgg cacgctgttg acgctgttaa gcgggtcgct gcagggtcgc tcggtgttcg
     5401 aggccacacg cgtcacctta atatgcgaag tggacctggg accgcgccgc cccgactgca
     5461 tctgcgtgtt cgaattcgcc aatgacaaga cgctgggcgg ggtttgtgtc atcatagaac
     5521 taaagacatg caaatatatt tcttccgggg acaccgccag caaacgcgag caacgggcca
     5581 cggggatgaa gcaggatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca
     5641 tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg
     5701 gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag
     5761 ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     5821 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc
     5881 agcccttgcg ccctgagtgc ttgcggcagc gtgaagcttg gcactggccg tcgttttaca
     5941 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     6001 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     6061 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     6121 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6181 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6241 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6301 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6361 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6421 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6481 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6541 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6601 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6661 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6721 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6781 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6841 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6901 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6961 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7021 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7201 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7501 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7561 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7621 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7801 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8161 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8221 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8281 gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
