LOCUS       sequence-220477-        5961 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 5961)
  AUTHORS   el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, 
            Lin D, Mercer WE, Kinzler KW, Vogelstein B
  TITLE     WAF1, a potential mediator of p53 tumor suppression.
  JOURNAL   Cell. 1993 Nov 19. 75(4):817-25.
  PUBMED    8242752
REFERENCE   2  (bases 1 to 5961)
  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: "Cell. 1993 
            Nov 19. 75(4):817-25."
COMMENT     Sequence Label: MG15-luc (mut p53 binding sites)
FEATURES             Location/Qualifiers
     source          1..5961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(410..426)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             480..2132
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(546..564)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          2263..2328
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2458..2478
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2903..3037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2994..3013
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3034..3050)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3087..3105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3126..3142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3126..3142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3139..3161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3150..3166
                     /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(3174..3180,3181..3198,3199..3204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3219..3240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3357..3374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3528..4116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3608..3627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4287..5078,5079..5147))
                     /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     4910..4929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5148..5252)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5278..5733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5415..5436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5627..5646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5860..5882
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5874..5891
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5875..5891
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5898..5917
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5898..5916
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5941..5957
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5942..5958
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcttgatcc ttaatggact ttaatggcct tatggacttt aatggcctta atggacttta
       61 atggccttaa tggactttaa tggccttaat ggactttaat ggccttaatg gactttaatg
      121 gccttaatgg actttaatgg ccttaatgga ctttaatggc cttaatggac tttaatggcc
      181 atcgaattcg atctgttttt tttagtatta agcagaggcc gggggcccct ggcctccgct
      241 tactctggag aaaaagaaga gaggcattcc agaggcaact tgtcaaaaca ggactggcgc
      301 cttggaggcg ctgtggggcc acccaaattg atataattaa gccccaaccg cctcttcccg
      361 cctcatttca gcctcaccac cctcgacaag cttatcgata ccgtcggggg atccactagt
      421 tctagagcgg ccgccaccgc ggtggagctc gagcttggca ttccggtact gttggtaaaa
      481 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatcctcta gaggatggaa
      541 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
      601 cttttacaga tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg
      661 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
      721 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
      781 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgaacattt
      841 cgcagcctac cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
      901 aaaaattacc aataatccag aaaattatta tcatggattc taaaacggat taccagggat
      961 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
     1021 ttgtaccaga gtcctttgat cgtgacaaaa caattgcact gataatgaat tcctctggat
     1081 ctactgggtt acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc
     1141 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
     1201 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
     1261 gagtcgtctt aatgtataga tttgaagaag agctgttttt acgatccctt caggattaca
     1321 aaattcaaag tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga
     1381 ttgacaaata cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga
     1441 aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg
     1501 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
     1561 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
     1621 cgctgggcgt taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt
     1681 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
     1741 gagacatagc ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt
     1801 taattaaata caaaggatat caggtggccc ccgctgaatt ggaatcgata ttgttacaac
     1861 accccaacat cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc
     1921 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
     1981 acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg
     2041 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
     2101 aggccaagaa gggcggaaag tccaaattgt aaaatgtaac tgtattcagc gatgacgaaa
     2161 ttcttagcta ttgtaatact ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg
     2221 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
     2281 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
     2341 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
     2401 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
     2461 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
     2521 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
     2581 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
     2641 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
     2701 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
     2761 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
     2821 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2881 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2941 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3001 gtccaaactc atcaatgtat cttatcatgt ctggatccac tagttctaga gcggccgcca
     3061 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
     3121 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3181 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     5281 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5341 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5401 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5461 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5521 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5581 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5641 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5701 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     5761 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     5821 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     5881 cgacggccag tgaattgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     5941 tcgaggtcga cggtatcgat a
//
