LOCUS       sequence-272183-        3391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2 copies of the p53 DNA binding motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3391)
  AUTHORS   Sarrion-Perdigones A, Gonzalez Y, Venken KJT
  TITLE     Rapid and Efficient Synthetic Assembly of Multiplex Luciferase 
            Reporter Plasmids for the Simultaneous Monitoring of Up to Six 
            Cellular Signaling Pathways.
  JOURNAL   Curr Protoc Mol Biol. 2020 Jun;131(1):e121. doi: 10.1002/cpmb.121.
  PUBMED    32539183
REFERENCE   2  (bases 1 to 3391)
  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: "Curr Protoc
            Mol Biol."; date: "2020-06"; volume: "131(1):e121. doi"; pages: " 
            10.1002/cpmb.121"
COMMENT     Sequence Label: TB:2xP53_RE:MiniP
FEATURES             Location/Qualifiers
     source          1..3391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        205..236
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      306..315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      310..319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(349..368)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(350..368)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(375..392)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(375..391)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(384..406)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      532..987
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(619..638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     829..850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1065..1169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1170..1238,1239..2030)
                     /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     complement(1388..1407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2201..2789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2690..2709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2943..2960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3077..3098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3113..3118,3119..3136,3137..3143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3151..3167
                     /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)."
     primer_bind     3156..3178
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3175..3191
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3175..3191
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3209..3227
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3209..3226
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    3330..3378
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctcgctgtca ggagtacaga acatgtctaa
      121 gcatgctgtg ccttgcctgg acttgcctgg ccttgccttc cctcgaggat atcaaatctg
      181 gcctcggcgg ccaagcttag acactagagg gtatataatg gaagctcgac ttccagcttg
      241 gcaatccggt actgttggta aagccaccaa tgcgagacca ctcatcgccc aatcgaattc
      301 ccgcggccgc catggcggcc gggagcatgc gacgtcgggc ccaattcgcc ctatagtgag
      361 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      421 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      481 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
      541 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      601 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      661 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      721 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccagcgccct
      781 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      841 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
      901 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      961 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     1021 cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1081 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1141 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1201 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1261 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1321 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1381 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1441 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1501 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1561 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1621 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1681 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1741 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1801 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1861 ataaatctgg agccggtgag cgtgggtcgc gcggtatcat tgcagcactg gggccagatg
     1921 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1981 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2041 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2101 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2161 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2221 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2281 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2341 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2401 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2461 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2521 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2581 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2641 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2701 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2761 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2821 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2881 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2941 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3001 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3061 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     3121 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     3181 acagctatga ccatgattac gccaagctat ttaggtgaca ctatagaata ctcaagctat
     3241 gcatccaacg cgttgggagc tctcccatat ggtcgacctg caggcggccg cgaattcact
     3301 agtgattggg cgatgagtgg tctcgggaga ataaaatatc tttattttca ttacatctgt
     3361 gtgttggttt tttgtgtgaa tcgatagtac t
//
