LOCUS       sequence-391296-        6838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human superstable p53 as His6GB1tev fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  AUTHORS   Krois AS, Park S, Martinez-Yamout MA, Dyson HJ, Wright PE
  TITLE     Mapping Interactions of the Intrinsically Disordered C-Terminal 
            Regions of Tetrameric p53 by Segmental Isotope Labeling and NMR.
  JOURNAL   Biochemistry. 2022 Dec 6;61(23):2709-2719. doi: 
            10.1021/acs.biochem.2c00528. Epub 2022 Nov 15.
  PUBMED    36380579
REFERENCE   2  (bases 1 to 6838)
  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"; doi: 
            "10.1021/acs.biochem.2c00528"; journalName: "Biochemistry"; date: 
            "2022-12-6- 6"; volume: "61"; issue: "23"; pages: "2709-2719"
COMMENT     Sequence Label: H6GB1tevP53SS(1-393)
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     197..216
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     300..319
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        300..318
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    319..343
                     /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)."
     RBS             358..380
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             393..410
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             426..587
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             612..632
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             639..1820
                     /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
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKLFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRAEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCYSSCMGGMNRRPILTIITLEDSSGNLLGRDSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     CDS             1869..1886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1939..1957)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1953..2000
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2037..2492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2124..2143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2334..2355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2518..2622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2623..2691,2692..3483)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2841..2860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3654..4242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4143..4162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4396..4413
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4428..4567
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4493..4512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4653..4675
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4669..4860)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5669..6751)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6713..6732
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6752..6829)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
       61 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
      121 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
      181 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
      241 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
      301 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
      361 gtttaacttt aagaaggaga tataccatgg gccatcatca tcatcatcac agcagcggca
      421 tgacttacaa actgatcctt aacggtaaaa ctttgaaagg cgaaaccact accgaagctg
      481 ttgatgctgc gactgcagaa aaagttttca aacagtacgc taacgacaac ggtgttgacg
      541 gtgaatggac ttacgacgat gcgactaaga ccttcaccgt tactgaagaa ttcagcagcg
      601 gcagcagcgg cgaaaacctg tattttcagg gctcccatat ggaggagccg cagtcagatc
      661 ctagcgtcga gccccctctg agtcaggaaa cattttcaga cctatggaaa ctacttcctg
      721 aaaacaacgt tctgtccccc ttgccgtccc aagcaatgga tgatttgatg ctgtccccgg
      781 acgatattga acaatggttc accgaagacc caggtccaga tgaagctccc agaatgccag
      841 aggctgctcc ccccgtggcc cctgcaccag cagctcctac accggcggcc cctgcaccag
      901 ccccctcctg gcccctgtca tcttctgtcc cttcccagaa aacctaccag ggcagctacg
      961 gtttccgtct gggcttcttg cattctggga cagccaagtc tgtgacttgc acgtactccc
     1021 ctgccctcaa caagctgttt tgccaactgg ccaagacctg ccctgtgcag ctgtgggttg
     1081 attccacacc cccgcccggc acccgcgtcc gcgccatggc catctacaag cagtcacagc
     1141 acatgacgga ggttgtgagg cgctgccccc accatgagcg ctgctcagat agcgatggtc
     1201 tggcccctcc tcagcatctt atccgagtgg aaggaaattt gcgtgcagag tatttggatg
     1261 acagaaacac ttttcgacat agtgtggtgg tgccctatga gccgcctgag gttggctctg
     1321 actgtaccac catccactac aactacatgt gttatagttc ctgcatgggc ggcatgaacc
     1381 ggaggcccat cctcaccatc atcacactgg aagactccag tggtaatcta ctgggacggg
     1441 atagctttga ggtgcgtgtt tgtgcctgtc ctgggagaga ccggcgcaca gaggaagaga
     1501 atctccgcaa gaaaggggag cctcaccacg agctgccccc agggagcact aagcgagcac
     1561 tgcccaacaa caccagctcc tctccccagc caaagaagaa accactggat ggagaatatt
     1621 tcacccttca gatccgtggg cgtgagcgct tcgagatgtt ccgagagctg aatgaggcct
     1681 tggaactcaa ggatgcccag gctgggaagg agccaggggg gagcagggct cactccagcc
     1741 acctgaagtc caaaaagggt cagtctacct cccgccataa aaaactcatg ttcaagacag
     1801 aagggcctga ctcagactaa taggatccga attcgagctc cgtcgacaag cttgcggccg
     1861 cactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     1921 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     1981 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     2041 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2101 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2161 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2221 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2281 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2341 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2401 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2461 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     2521 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2581 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2641 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2701 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2761 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2821 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2881 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2941 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3001 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3061 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3121 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     3181 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3241 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3301 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3361 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3421 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3481 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3541 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3601 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3661 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3721 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3781 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3841 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3901 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3961 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4021 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4081 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4141 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4201 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4261 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4321 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4381 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4441 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     4501 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4561 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4621 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4681 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4741 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4801 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4861 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     4921 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     4981 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     5041 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     5101 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     5161 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     5221 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     5281 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     5341 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     5401 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     5461 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     5521 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     5581 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     5641 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5761 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     5821 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     5881 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     5941 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     6001 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     6061 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     6121 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     6181 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     6241 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     6301 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     6361 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     6421 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     6481 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     6541 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     6601 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     6661 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     6721 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     6781 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctc
//
