LOCUS       sequence-197539-        6878 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 6878)
  AUTHORS   Ayed A, Mulder FA, Yi GS, Lu Y, Kay LE, Arrowsmith CH
  TITLE     Latent and active p53 are identical in conformation.
  JOURNAL   Nat Struct Biol. 2001 Sep . 8(9):756-60.
  PUBMED    11524676
REFERENCE   2  (bases 1 to 6878)
  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: "Nat Struct 
            Biol. 2001 Sep . 8(9):756-60."
COMMENT     Sequence Label: human p53-(1-393)
FEATURES             Location/Qualifiers
     source          1..6878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(325..1506)
                     /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"
     CDS             complement(1510..1527)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1537..1554)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1573..1595
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1610..1634
                     /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     complement(1634..1653)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1635..1653)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1737..1756)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1962..2039
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(2059..2078)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4433..4624
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4618..4640)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4726..4865
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4781..4800
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4880..4897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5051..5639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5131..5150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5810..6601,6602..6670))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6433..6452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6671..6775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6843..6861
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctcagtc tgagtcaggc ccttctgtct tgaacatgag
      361 ttttttatgg cgggaggtag actgaccctt tttggacttc aggtggctgg agtgagccct
      421 gctcccccct ggctccttcc cagcctgggc atccttgagt tccaaggcct cattcagctc
      481 tcggaacatc tcgaagcgct cacgcccacg gatctgaagg gtgaaatatt ctccatccag
      541 tggtttcttc tttggctggg gagaggagct ggtgttgttg ggcagtgctc gcttagtgct
      601 ccctgggggc agctcgtggt gaggctcccc tttcttgcgg agattctctt cctctgtgcg
      661 ccggtctctc ccaggacagg cacaaacacg cacctcaaag ctgttccgtc ccagtagatt
      721 accactggag tcttccagtg tgatgatggt gaggatgggc ctccggttca tgccgcccat
      781 gcaggaactg ttacacatgt agttgtagtg gatggtggta cagtcagagc caacctcagg
      841 cggctcatag ggcaccacca cactatgtcg aaaagtgttt ctgtcatcca aatactccac
      901 acgcaaattt ccttccactc ggataagatg ctgaggaggg gccagaccat cgctatctga
      961 gcagcgctca tggtgggggc agcgcctcac aacctccgtc atgtgctgtg actgcttgta
     1021 gatggccatg gcgcggacgc gggtgccggg cgggggtgtg gaatcaaccc acagctgcac
     1081 agggcaggtc ttggccagtt ggcaaaacat cttgttgagg gcaggggagt acgtgcaagt
     1141 cacagacttg gctgtcccag aatgcaagaa gcccagacgg aaaccgtagc tgccctggta
     1201 ggttttctgg gaagggacag aagatgacag gggccaggag ggggctggtg caggggccgc
     1261 cggtgtagga gctgctggtg caggggccac ggggggagca gcctctggca ttctgggagc
     1321 ttcatctgga cctgggtctt cggtgaacca ttgttcaata tcgtccgggg acagcatcaa
     1381 atcatccatt gcttgggacg gcaaggggga cagaacgttg ttttcaggaa gtagtttcca
     1441 taggtctgaa aatgtttcct gactcagagg gggctcgacg ctaggatctg actgcggctc
     1501 ctccatatgg ctgccgcgcg gcaccaggcc gctgctgtga tgatgatgat gatggctgct
     1561 gcccatggta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
     1621 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct
     1681 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     1741 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     1801 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     1861 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     1921 cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca
     1981 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     2041 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     2101 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     2161 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     2221 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     2281 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     2341 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     2401 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     2461 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     2521 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     2581 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     2641 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     2701 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     2761 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     2821 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
     2881 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     2941 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     3001 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     3061 cgttggccga ttcattaatg cagctggcac gacagtttcc cgactggaaa gcgggcagtg
     3121 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc
     3181 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     3241 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     3301 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     3361 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     3421 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     3481 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     3541 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     3601 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc
     3661 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     3721 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     3781 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     3841 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     3901 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     3961 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     4021 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     4081 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     4141 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     4201 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     4261 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     4321 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     4381 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     4441 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     4501 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     4561 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     4621 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4681 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     4741 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     4801 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4861 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5461 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6121 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6421 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6781 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6841 aaaataggcg tatcacgagg ccctttcgtc ttcaagaa
//
