LOCUS       sequence-139148-        6869 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 6869)
  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 6869)
  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 wt/EGFP
FEATURES             Location/Qualifiers
     source          1..6869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     370..389
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    388..443
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     489..508
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          644..1216
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             complement(1785..2966)
                     /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"
     primer_bind     complement(3044..3060)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3045..3061)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3201..3239)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    3272..3290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3314..3332
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3356..3374
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3398..3416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3440..3458
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3601..3617
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3601..3617
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(3625..3627,3628..3630,3631..4344)
                     /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             join(3625..3627,3628..3630,3631..4341)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3670..3691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3931..3950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4278..4299
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4381..4515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4405..4424)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4459..4478
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4878..4896)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4964..5068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5069..5137,5138..5929)
                     /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     complement(5287..5306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6100..6688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6589..6608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6842..6859
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagtcgag
       61 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
      121 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
      181 acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata aggaatggac
      241 agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt tcatggcagc
      301 cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat gttttaaatg
      361 atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac acactattgc
      421 aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct tcatgatgtc
      481 cccataattt ttggcagagg gaaaaagatc gatctcagtg gtatttgtga gccagggcat
      541 tggccacacc agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattctttgc
      601 caaaatgatg agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga
      661 aggcatgaac atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct
      721 caaacagtgt atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt
      781 tgtttctacc aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac
      841 cttatcattt aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac
      901 cctgattgcc ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa
      961 aagtgaattt aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa
     1021 atgaaaagaa aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa
     1081 attatcatga gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga
     1141 aaactttgcc ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa
     1201 gggtccccaa actcaccctg aagttctcag ctctagagat atcgtcgagg ccgcgctagc
     1261 gtttaaactg tcgaccttaa ttaagagatc ccgctagacc ccatgtaata aaaggtggtt
     1321 tcaaggccag atgtacatta tttcattaac cctcacaatg cactctgtga ggtaggtgca
     1381 aatgccagca tttcacagat atgggccttg aagttagaga aaattcaaca gtgagggaca
     1441 gcttccctgg ttagtacggt gaagtgggcc cctacctaga atgtggctga ttgtaaacta
     1501 acccttaact gcaagaacat ttcttacatc tcccaaacat ccctcacagt aaaaacctta
     1561 aaatctaagc tggtatgtcc tactccccat cctcctcccc acaacaaaac accagtgcag
     1621 gccaacttgt tcagtggagc cccgggacaa agcaaatgga agtcctgggt gcttctgacg
     1681 cacacctatt gcaagcaagg gttcaaagac ccaaaaccca aaatggcagg ggagggagag
     1741 atgggggtgg gaggctgtca gtggggaaca agaagtggag aatgtcagtc tgagtcaggc
     1801 ccttctgtct tgaacatgag ttttttatgg cgggaggtag actgaccctt tttggacttc
     1861 aggtggctgg agtgagccct gctcccccct ggctccttcc cagcctgggc atccttgagt
     1921 tccaaggcct cattcagctc tcggaacatc tcgaagcgct cacgcccacg gatctgaagg
     1981 gtgaaatatt ctccatccag tggtttcttc tttggctggg gagaggagct ggtgttgttg
     2041 ggcagtgctc gcttagtgct ccctgggggc agctcgtggt gaggctcccc tttcttgcgg
     2101 agattctctt cctctgtgcg ccggtctctc ccaggacagg cacaaacacg cacctcaaag
     2161 ctgttccgtc ccagtagatt accactggag tcttccagtg tgatgatggt gaggatgggc
     2221 ctccggttca tgccgcccat gcaggaactg ttacacatgt agttgtagtg gatggtggta
     2281 cagtcagagc caacctcagg cggctcatag ggcaccacca cactatgtcg aaaagtgttt
     2341 ctgtcatcca aatactccac acgcaaattt ccttccactc ggataagatg ctgaggaggg
     2401 gccagaccat cgctatctga gcagcgctca tggtgggggc agcgcctcac aacctccgtc
     2461 atgtgctgtg actgcttgta gatggccatg gcgcggacgc gggtgccggg cgggggtgtg
     2521 gaatcaaccc acagctgcac agggcaggtc ttggccagtt ggcaaaacat cttgttgagg
     2581 gcaggggagt acgtgcaagt cacagacttg gctgtcccag aatgcaagaa gcccagacgg
     2641 aaaccgtagc tgccctggta ggttttctgg gaagggacag aagatgacag gggccaggag
     2701 ggggctggtg caggggccgc cggtgtagga gctgctggtg caggggccac gcggggagca
     2761 gcctctggca ttctgggagc ttcatctgga cctgggtctt cagtgaacca ttgttcaata
     2821 tcgtccgggg acagcatcaa atcatccatt gcttgggacg gcaaggggga cagaacgttg
     2881 ttttcaggaa gtagtttcca taggtctgaa aatgtttcct gactcagagg gggctcgacg
     2941 ctaggatctg actgcggctc ctccatggca gtgacccgga aggcagtctg gctgccaatc
     3001 cagggaagcg tgtcaccgtc gtggaattcg atatcaagct tatcgatacc gtcgacctcg
     3061 aggcgcgtca gctgactaga ggatccccgg gtaccgagct cgaattcggg gccgcggagg
     3121 ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg tctccaggcg
     3181 atctgacggt tcactaaacg agctctgctt atataggcct cccaccgtac acgcctacct
     3241 cgacccgggt accgagctcg actttcactt ttctctatca ctgataggga gtggtaaact
     3301 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct
     3361 atcactgata gggagtggta aactcgactt tcacttttct ctatcactga tagggagtgg
     3421 taaactcgac tttcactttt ctctatcact gatagggagt ggtaaactcg acctatataa
     3481 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     3541 tccatagaag acaccgggac cgatccagcc tccgcggccc cgaattcctg cagcggccgc
     3601 tctagaacta gtggatcccc gcggatggtg agcaagggcg aggagctgtt caccggggtg
     3661 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3721 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3781 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3841 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3901 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3961 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     4021 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     4081 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     4141 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4201 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     4261 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     4321 ggcatggacg agctgtacaa gtaagaattc gatatcaagc ttgtcgacct agttctagag
     4381 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4441 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4501 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4561 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     4621 cctgcaagcc tcgtcgtcct ggccggacca cgctatctgt gcaaggtccc cggccccgga
     4681 cgcgcgctcc atgagcagag cgcccgccgc cgaggcgaag actcgggcgg cgccctgccc
     4741 gtcccaccag gtcaacaggc ggtaaccggc ctcttcatcg ggaatgcgcg cgaccttcag
     4801 catcgccggc atgtccccct ggcggacggg aagtatccag ctcgaccaag ctggccgcgt
     4861 cgagtgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4921 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4981 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5041 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5101 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5161 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5221 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5281 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     5341 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5401 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5461 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5521 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5581 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5641 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5701 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5761 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5821 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5881 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5941 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6001 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6061 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6121 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6181 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6241 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6301 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6361 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6421 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6481 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6541 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6601 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6661 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6721 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6781 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6841 cagcgagtca gtgagcgagg aagcggaag
//
