LOCUS       sequence-154449-        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 R248W
FEATURES             Location/Qualifiers
     source          1..8318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        227..606
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        607..810
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     761..781
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     807..831
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          922..1494
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1696..2877
                     /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
                     PEVGSDCTTIHYNYMCNSSCMGGMNWRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(3466..3485)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3531..3586
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3585..3604)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3944..3960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3944..3960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3957..3979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3968..3984
                     /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)."
     promoter        complement(join(3992..3998,3999..4016,4017..4022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4037..4058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4175..4192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4346..4934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4426..4445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5105..5896,5897..5965))
                     /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     5728..5747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5966..6070)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6138..6156
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6194..6216)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6316..6335
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6529..6551
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6543..6560
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6544..6560
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6971..7116
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             7173..7967
                     /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(7227..7246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7837..7856
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8161..8180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8205..8252
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
       61 ggctcgcagc caacgtcggg gcggcaagcc ctgccatagc cacgggcccc gtgggttagg
      121 gacggcggat cgcggccctc tagagagctt ggcccattgc atacgttgta tccatatcat
      181 aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg
      241 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      301 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      361 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      421 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      481 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      541 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      601 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      661 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      721 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      781 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      841 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggtcg
      901 accgatcctg agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct
      961 attgtaaaat tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga
     1021 agatgtccct tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc
     1081 tgttgacaac cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac
     1141 tttagcttgc atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag
     1201 tactttctct aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca
     1261 cagttttaga gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat
     1321 tctggctggc gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg
     1381 cctttctctt tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag
     1441 tccaaaccgg gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg
     1501 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattcct cgacggatcc
     1561 cgctagagcc accgtccagg gagcaggtag ctgctgggct ccggggacac tttgcgttcg
     1621 ggctgggagc gtgctttcca cgacggtgac acgcttccct ggattggcag ccagactgcc
     1681 ttccgggtca ctgccatgga ggagccgcag tcagatccta gcgtcgagcc ccctctgagt
     1741 caggaaacat tttcagacct atggaaacta cttcctgaaa acaacgttct gtcccccttg
     1801 ccgtcccaag caatggatga tttgatgctg tccccggacg atattgaaca atggttcact
     1861 gaagacccag gtccagatga agctcccaga atgccagagg ctgctccccc cgtggcccct
     1921 gcaccagcag ctcctacacc ggcggcccct gcaccagccc cctcctggcc cctgtcatct
     1981 tctgtccctt cccagaaaac ctaccagggc agctacggtt tccgtctggg cttcttgcat
     2041 tctgggacag ccaagtctgt gacttgcacg tactcccctg ccctcaacaa gatgttttgc
     2101 caactggcca agacctgccc tgtgcagctg tgggttgatt ccacaccccc gcccggcacc
     2161 cgcgtccgcg ccatggccat ctacaagcag tcacagcaca tgacggaggt tgtgaggcgc
     2221 tgcccccacc atgagcgctg ctcagatagc gatggtctgg cccctcctca gcatcttatc
     2281 cgagtggaag gaaatttgcg tgtggagtat ttggatgaca gaaacacttt tcgacatagt
     2341 gtggtggtgc cctatgagcc gcctgaggtt ggctctgact gtaccaccat ccactacaac
     2401 tacatgtgta acagttcctg catgggcggc atgaactgga ggcccatcct caccatcatc
     2461 acactggaag actccagtgg taatctactg ggacggaaca gctttgaggt gcgtgtttgt
     2521 gcctgtcctg ggagagaccg gcgcacagag gaagagaatc tccgcaagaa aggggagcct
     2581 caccacgagc tgcccccagg gagcactaag cgagcactgc ccaacaacac cagctcctct
     2641 ccccagccaa agaagaaacc actggatgga gaatatttca cccttcagat ccgtgggcgt
     2701 gagcgcttcg agatgttccg agagctgaat gaggccttgg aactcaagga tgcccaggct
     2761 gggaaggagc caggggggag cagggctcac tccagccacc tgaagtccaa aaagggtcag
     2821 tctacctccc gccataaaaa actcatgttc aagacagaag ggcctgactc agactgacat
     2881 tctccacttc ttgttcccca ctgacagcct cccaccccca tctctccctc ccctgccatt
     2941 ttgggttttg ggtctttgaa cccttgcttg caataggtgt gcgtcagaag cacccaggac
     3001 ttccatttgc tttgtcccgg ggctccactg aacaagttgg cctgcactgg tgttttgttg
     3061 tggggaggag gatggggagt aggacatacc agcttagatt ttaaggtttt tactgtgagg
     3121 gatgtttggg agatgtaaga aatgttcttg cagttaaggg ttagtttaca atcagccaca
     3181 ttctaggtag gggcccactt caccgtacta accagggaag ctgtccctca ctgttgaatt
     3241 ttctctaact tcaaggccca tatctgtgaa atgctggcat ttgcacctac ctcacagagt
     3301 gcattgtgag ggttaatgaa ataatgtaca tctggccttg aaaccacctt ttattacatg
     3361 gggtctagcg ggatccgtcg aggaattcac tcctcaggtg caggctgcct atcagaaggt
     3421 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     3481 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     3541 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3601 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     3661 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     3721 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     3781 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     3841 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     3901 cttatggaga tccctcgacg gatcggccgc aattcgtaat catgtcatag ctgtttcctg
     3961 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4021 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4081 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4141 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4981 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5041 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5101 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5161 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5221 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5281 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5341 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5401 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5461 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5521 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5581 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5641 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5701 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5761 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5821 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5881 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5941 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6001 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6061 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6121 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6181 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6241 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6301 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6361 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     6421 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     6481 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     6541 gttgtaaaac gacggccagt gccaagcttc acgctgccgc aagcactcag ggcgcaaggg
     6601 ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat
     6661 gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt
     6721 agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga
     6781 accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg
     6841 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatcct gcttcatccc
     6901 cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt
     6961 tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca
     7021 gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc
     7081 ctcgaacacc gagcgaccct gcagcgaccc gcttaacagc gtcaacagcg tgccgcagat
     7141 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     7201 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     7261 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     7321 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     7381 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     7441 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     7501 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     7561 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     7621 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     7681 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     7741 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     7801 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     7861 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     7921 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     7981 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     8041 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     8101 ccagcgcggg gatctcatgc tggagttctt cgcccacccc gggagatggg ggaggctaac
     8161 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     8221 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     8281 gcactctgtc gataccccac cgagacccca ttggggcc
//
