LOCUS       sequence-208221-        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 wt
FEATURES             Location/Qualifiers
     source          1..8318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1470..2651
                     /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
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(3240..3259)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3305..3360
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3359..3378)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3718..3734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3718..3734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3731..3753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3742..3758
                     /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(3766..3772,3773..3790,3791..3796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3811..3832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3949..3966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4120..4708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4200..4219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4879..5670,5671..5739))
                     /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     5502..5521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5740..5844)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5912..5930
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5968..5990)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6090..6109
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6303..6325
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6317..6334
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6318..6334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6745..6890
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6947..7741
                     /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(7001..7020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7611..7630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7935..7954)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7979..8026
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atcccgctag agccaccgtc cagggagcag gtagctgctg ggctccgggg
     1381 acactttgcg ttcgggctgg gagcgtgctt tccacgacgg tgacacgctt ccctggattg
     1441 gcagccagac tgccttccgg gtcactgcca tggaggagcc gcagtcagat cctagcgtcg
     1501 agccccctct gagtcaggaa acattttcag acctatggaa actacttcct gaaaacaacg
     1561 ttctgtcccc cttgccgtcc caagcaatgg atgatttgat gctgtccccg gacgatattg
     1621 aacaatggtt cactgaagac ccaggtccag atgaagctcc cagaatgcca gaggctgctc
     1681 cccccgtggc ccctgcacca gcagctccta caccggcggc ccctgcacca gccccctcct
     1741 ggcccctgtc atcttctgtc ccttcccaga aaacctacca gggcagctac ggtttccgtc
     1801 tgggcttctt gcattctggg acagccaagt ctgtgacttg cacgtactcc cctgccctca
     1861 acaagatgtt ttgccaactg gccaagacct gccctgtgca gctgtgggtt gattccacac
     1921 ccccgcccgg cacccgcgtc cgcgccatgg ccatctacaa gcagtcacag cacatgacgg
     1981 aggttgtgag gcgctgcccc caccatgagc gctgctcaga tagcgatggt ctggcccctc
     2041 ctcagcatct tatccgagtg gaaggaaatt tgcgtgtgga gtatttggat gacagaaaca
     2101 cttttcgaca tagtgtggtg gtgccctatg agccgcctga ggttggctct gactgtacca
     2161 ccatccacta caactacatg tgtaacagtt cctgcatggg cggcatgaac cggaggccca
     2221 tcctcaccat catcacactg gaagactcca gtggtaatct actgggacgg aacagctttg
     2281 aggtgcgtgt ttgtgcctgt cctgggagag accggcgcac agaggaagag aatctccgca
     2341 agaaagggga gcctcaccac gagctgcccc cagggagcac taagcgagca ctgcccaaca
     2401 acaccagctc ctctccccag ccaaagaaga aaccactgga tggagaatat ttcacccttc
     2461 agatccgtgg gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc ttggaactca
     2521 aggatgccca ggctgggaag gagccagggg ggagcagggc tcactccagc cacctgaagt
     2581 ccaaaaaggg tcagtctacc tcccgccata aaaaactcat gttcaagaca gaagggcctg
     2641 actcagactg acattctcca cttcttgttc cccactgaca gcctcccacc cccatctctc
     2701 cctcccctgc cattttgggt tttgggtctt tgaacccttg cttgcaatag gtgtgcgtca
     2761 gaagcaccca ggacttccat ttgctttgtc ccggggctcc actgaacaag ttggcctgca
     2821 ctggtgtttt gttgtgggga ggaggatggg gagtaggaca taccagctta gattttaagg
     2881 tttttactgt gagggatgtt tgggagatgt aagaaatgtt cttgcagtta agggttagtt
     2941 tacaatcagc cacattctag gtaggggccc acttcaccgt actaaccagg gaagctgtcc
     3001 ctcactgttg aattttctct aacttcaagg cccatatctg tgaaatgctg gcatttgcac
     3061 ctacctcaca gagtgcattg tgagggttaa tgaaataatg tacatctggc cttgaaacca
     3121 ccttttatta catggggtct agcgggatcc gtcgaggaat tcactcctca ggtgcaggct
     3181 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     3241 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     3301 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     3361 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     3421 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     3481 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     3541 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     3601 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     3661 gctgtccctc ttctcttatg gagatccctc gacggatcgg ccgcaattcg taatcatgtc
     3721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4981 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5401 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5761 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5821 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5881 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5941 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6001 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6061 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6121 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6181 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6241 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6301 ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttcacgctg ccgcaagcac
     6361 tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg
     6421 tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca
     6481 aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta
     6541 tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc
     6601 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga
     6661 tcctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata
     6721 tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg
     6781 aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc gcatattaag
     6841 gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa cagcgtcaac
     6901 agcgtgccgc agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     6961 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     7021 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     7081 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     7141 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     7201 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     7261 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     7321 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     7381 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     7441 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     7501 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     7561 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     7621 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     7681 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     7741 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     7801 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     7861 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggaga
     7921 tgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     7981 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     8041 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     8101 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     8161 gccaacgtcg gggcggcaag ccctgccata gccacgggcc ccgtgggtta gggacggcgg
     8221 atcgcggccc tctagagagc ttggcccatt gcatacgttg tatccatatc ataatatgta
     8281 catttatatt ggctcatgtc caacattacc gccatgtt
//
