LOCUS       sequence-251850-        6459 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 6459)
  AUTHORS   Yu J, Zhang L, Hwang PM, Rago C, Kinzler KW, Vogelstein B
  TITLE     Identification and classification of p53-regulated genes.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Dec 7. 96(25):14517-22.
  PUBMED    10588737
REFERENCE   2  (bases 1 to 6459)
  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: "Proc Natl 
            Acad Sci U S A. 1999 Dec 7. 96(25):14517-22."
COMMENT     Sequence Label: pBI-p53 R175H/EGFP
FEATURES             Location/Qualifiers
     source          1..6459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    84..102
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    126..144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    168..186
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    210..228
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    252..270
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        303..341
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     481..497
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     482..498
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             576..1757
                     /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
                     KQSQHMTEVVRHCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     intron          1916..2488
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2624..2643)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2689..2744
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2743..2762)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3142..3159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3313..3901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3393..3412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4072..4863,4864..4932))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4695..4714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4933..5037)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5105..5123
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    5486..5620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5523..5542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5577..5596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5657..6370,6371..6373,6374..6376))
                     /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             complement(join(5660..6370,6371..6373,6374..6376))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5702..5723)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6051..6070
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6310..6331
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6384..6400)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6384..6400)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 aggtcaaaac agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg
       61 cttatatagg tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
      121 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
      181 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
      241 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg tacccgggtc
      301 gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg aaccgtcaga
      361 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      421 gcctccgcgg ccccgaattc gagctcggta cccggggatc ctctagtcag ctgacgcgcc
      481 tcgaggtcga cggtatcgat aagcttgata tcgaattcca cgacggtgac acgcttccct
      541 ggattggcag ccagactgcc ttccgggtca ctgccatgga ggagccgcag tcagatccta
      601 gcgtcgagcc ccctctgagt caggaaacat tttcagacct atggaaacta cttcctgaaa
      661 acaacgttct gtcccccttg ccgtcccaag caatggatga tttgatgctg tccccggacg
      721 atattgaaca atggttcact gaagacccag gtccagatga agctcccaga atgccagagg
      781 ctgctccccg cgtggcccct gcaccagcag ctcctacacc ggcggcccct gcaccagccc
      841 cctcctggcc cctgtcatct tctgtccctt cccagaaaac ctaccagggc agctacggtt
      901 tccgtctggg cttcttgcat tctgggacag ccaagtctgt gacttgcacg tactcccctg
      961 ccctcaacaa gatgttttgc caactggcca agacctgccc tgtgcagctg tgggttgatt
     1021 ccacaccccc gcccggcacc cgcgtccgcg ccatggccat ctacaagcag tcacagcaca
     1081 tgacggaggt tgtgaggcac tgcccccacc atgagcgctg ctcagatagc gatggtctgg
     1141 cccctcctca gcatcttatc cgagtggaag gaaatttgcg tgtggagtat ttggatgaca
     1201 gaaacacttt tcgacatagt gtggtggtgc cctatgagcc gcctgaggtt ggctctgact
     1261 gtaccaccat ccactacaac tacatgtgta acagttcctg catgggcggc atgaaccgga
     1321 ggcccatcct caccatcatc acactggaag actccagtgg taatctactg ggacggaaca
     1381 gctttgaggt gcgtgtttgt gcctgtcctg ggagagaccg gcgcacagag gaagagaatc
     1441 tccgcaagaa aggggagcct caccacgagc tgcccccagg gagcactaag cgagcactgc
     1501 ccaacaacac cagctcctct ccccagccaa agaagaaacc actggatgga gaatatttca
     1561 cccttcagat ccgtgggcgt gagcgcttcg agatgttccg agagctgaat gaggccttgg
     1621 aactcaagga tgcccaggct gggaaggagc cgggggggag cagggctcac tccagccacc
     1681 tgaagtccaa aaagggtcag tctacctccc gccataaaaa actcatgttc aagacagaag
     1741 ggcctgactc agactgacat tctccacttc ttgttcccca ctgacagcct cccaccccca
     1801 tctctccctc ccctgccatt ttgaattcct gcagcccggg ggatctctta attaaggtcg
     1861 acagtttaaa cgctagcgcg gcctcgacga tatctctaga gctgagaact tcagggtgag
     1921 tttggggacc cttgattgtt ctttcttttt cgctattgta aaattcatgt tatatggagg
     1981 gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt cccttgtatc accatggacc
     2041 ctcatgataa ttttgtttct ttcactttct actctgttga caaccattgt ctcctcttat
     2101 tttcttttca ttttctgtaa ctttttcgtt aaactttagc ttgcatttgt aacgaatttt
     2161 taaattcact tttgtttatt tgtcagattg taagtacttt ctctaatcac ttttttttca
     2221 aggcaatcag ggtatattat attgtacttc agcacagttt tagagaacaa ttgttataat
     2281 taaatgataa ggtagaatat ttctgcatat aaattctggc tggcgtggaa atattcttat
     2341 tggtagaaac aactacaccc tggtcatcat cctgcctttc tctttatggt tacaatgata
     2401 tacactgttt gagatgagga taaaatactc tgagtccaaa ccgggcccct ctgctaacca
     2461 tgttcatgcc ttcttctctt tcctacagct cctgggcaac gtgctggttg ttgtgctgtc
     2521 tcatcatttt ggcaaagaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc
     2581 tggtgtggcc aatgccctgg ctcacaaata ccactgagat cgatcttttt ccctctgcca
     2641 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     2701 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     2761 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     2821 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     2881 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     2941 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     3001 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     3061 tatggagatc cctcgactgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3121 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3181 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3241 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3301 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3361 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3421 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3481 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3541 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3601 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3661 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3721 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3781 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3841 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3901 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3961 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4021 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4081 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4141 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4201 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4261 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4321 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4381 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4441 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4501 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4561 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4621 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4681 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4741 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4801 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4861 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4921 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4981 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5041 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5101 taaaaatagg cgtatcacga ggccctttcg tcttcactcg acgcggccag cttggtcgag
     5161 ctggatactt cccgtccgcc agggggacat gccggcgatg ctgaaggtcg cgcgcattcc
     5221 cgatgaagag gccggttacc gcctgttgac ctggtgggac gggcagggcg ccgcccgagt
     5281 cttcgcctcg gcggcgggcg ctctgctcat ggagcgcgcg tccggggccg gggaccttgc
     5341 acagatagcg tggtccggcc aggacgacga ggcttgcagg atcataatca gccataccac
     5401 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     5461 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     5521 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5581 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc ctctagaact aggtcgacaa
     5641 gcttgatatc gaattcttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     5701 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     5761 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     5821 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     5881 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     5941 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     6001 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     6061 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     6121 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     6181 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     6241 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     6301 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     6361 cgcccttgct caccatccgc ggggatccac tagttctaga gcggccgctg caggaattcg
     6421 gggccgcgga ggctggatcg gtcccggtgt cttctatgg
//
