LOCUS       sequence-433635-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of P53 NTAD-DBD for segmental labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  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 7130)
  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: H6GB1tevP53(1-303)IntN
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..288
                     /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             313..333
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1862..1879
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1932..1950)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1946..1993
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2030..2485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2117..2136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2327..2348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2511..2615
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2616..2684,2685..3476)
                     /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(2834..2853)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3647..4235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4136..4155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4389..4406
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4421..4560
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4486..4505)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4646..4668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4662..4853)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5662..6744)
                     /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     6706..6725
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6745..6822)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7028..7047
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggccatcatc atcatcatca cagcagcggc
      121 atgacttaca aactgatcct taacggtaaa actttgaaag gcgaaaccac taccgaagct
      181 gttgatgctg cgactgcaga aaaagttttc aaacagtacg ctaacgacaa cggtgttgac
      241 ggtgaatgga cttacgacga tgcgactaag accttcaccg ttactgaaga attcagcagc
      301 ggcagcagcg gcgaaaacct gtattttcag ggctcccata tggaggagcc gcagtcagat
      361 cctagcgtcg agccccctct gagtcaggaa acattttcag acctatggaa actacttcct
      421 gaaaacaacg ttctgtcccc cttgccgtcc caagcaatgg atgatttgat gctgtccccg
      481 gacgatattg aacaatggtt caccgaagac ccaggtccag atgaagctcc cagaatgcca
      541 gaggctgctc cccccgtggc ccctgcacca gcagctccta caccggcggc ccctgcacca
      601 gccccctcct ggcccctgtc atcttctgtc ccttcccaga aaacctacca gggcagctac
      661 ggtttccgtc tgggcttctt gcattctggg acagccaagt ctgtgacttg cacgtactcc
      721 cctgccctca acaagctgtt ttgccaactg gccaagacct gccctgtgca gctgtgggtt
      781 gattccacac ccccgcccgg cacccgcgtc cgcgccatgg ccatctacaa gcagtcacag
      841 cacatgacgg aggttgtgag gcgctgcccc caccatgagc gctgctcaga tagcgatggt
      901 ctggcccctc ctcagcatct tatccgagtg gaaggaaatt tgcgtgcaga gtatttggat
      961 gacagaaaca cttttcgaca tagtgtggtg gtgccctatg agccgcctga ggttggctct
     1021 gactgtacca ccatccacta caactacatg tgttatagtt cctgcatggg cggcatgaac
     1081 cggaggccca tcctcaccat catcacactg gaagactcca gtggtaatct actgggacgg
     1141 gatagctttg aggtgcgtgt ttgtgcctgt cctgggagag accggcgcac agaggaagag
     1201 aatctccgca agaaagggga gcctcaccac gagctgcccc caggatcctg tttaagctat
     1261 gaaacggaaa tattgacagt agaatatgga ttattaccga ttggtaaaat tgtagaaaag
     1321 cgcatcgaat gtactgttta tagcgttgat aataatggaa atatttatac acaacctgta
     1381 gcacaatggc acgatcgcgg agaacaagag gtgtttgagt attgtttgga agatggttca
     1441 ttgattcggg caacaaaaga ccataagttt atgactgttg atggtcaaat gttgccaatt
     1501 gatgaaatat ttgaacgtga attggatttg atgcgggttg ataatttgcc gaatatcaaa
     1561 atagccacac gtaaatattt aggcaaacaa aatgtctatg acattggagt tgagcgctaa
     1621 cataattttg cactcaaaaa tggcttcata gcttctaatt gtttcaatgg tacccagtac
     1681 aaacttatcc tgaacggtaa aaccctgaaa ggtgaaacca ccaccgaagc tgttgacgct
     1741 gctaccgcgg aaaaagtttt caaacagtac gctaacgaca acggtgttga cggtgaatgg
     1801 acctacgacg acgctaccaa aacctacacc gtaacggaat aagcttgcgg ccgcactcga
     1861 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     1921 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1981 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     2041 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2101 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2161 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2221 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2281 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2341 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2401 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2461 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2521 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2581 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2641 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2701 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2761 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2821 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2881 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2941 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3001 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3061 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3121 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     3181 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3241 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3301 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3361 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3421 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3481 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3541 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3601 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3661 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3721 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3781 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3841 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3901 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3961 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4021 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4081 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4141 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4201 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4261 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4321 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4381 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4441 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     4501 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     4561 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4621 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4681 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     4741 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     4801 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     4861 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     4921 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     4981 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     5041 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     5101 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     5161 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     5221 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     5281 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     5341 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     5401 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     5461 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     5521 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     5581 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     5641 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5701 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     5761 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     5821 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     5881 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     5941 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     6001 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     6061 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     6121 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     6181 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     6241 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     6301 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     6361 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     6421 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     6481 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     6541 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     6601 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     6661 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     6721 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     6781 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     6841 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     6901 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     6961 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     7021 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     7081 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
