LOCUS       sequence-398143-        5905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of p53 TET-CTD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5905)
  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 5905)
  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(304-393)
FEATURES             Location/Qualifiers
     source          1..5905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             144..164
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             468..485
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(538..556)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      552..599
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      636..1091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(723..742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     933..954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1117..1221
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1222..1290,1291..2082)
                     /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(1440..1459)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2253..2841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2742..2761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2995..3012
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3027..3166
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3092..3111)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3252..3274
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3268..3459)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4268..5350)
                     /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
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITEYGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5312..5331
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5351..5428)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5634..5653
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5737..5756
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5737..5755
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5756..5780
                     /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             5795..5817
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5830..5847
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(5863..5905,1..119)
                     /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"
ORIGIN
        1 ctaccgaagc tgttgatgct gcgactgcag aaaaagtttt caaacagtac gctaacgaca
       61 acggtgttga cggtgaatgg acttacgacg atgcgactaa gaccttcacc gttactgaag
      121 aattcagcag cggcagcagc ggcgaaaacc tgtattttca gggctcccat actaagcgag
      181 cactgcccaa caacaccagc tcctctcccc agccaaagaa gaaaccactg gatggagaat
      241 atttcaccct tcagatccgt gggcgtgagc gcttcgagat gttccgagag ctgaatgagg
      301 ccttggaact caaggatgcc caggctggga aggagccagg ggggagcagg gctcactcca
      361 gccacctgaa gtccaaaaag ggtcagtcta cctcccgcca taaaaaactc atgttcaaga
      421 cagaagggcc tgactcagac taatagaagc ttgcggccgc actcgagcac caccaccacc
      481 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      541 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      601 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
      661 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      721 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccatcaagc
      781 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      841 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      901 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
      961 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1021 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1081 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1141 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1201 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1261 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1321 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1381 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1441 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1501 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1561 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1621 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1681 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1741 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     1801 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1861 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1921 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1981 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2041 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2101 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2161 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2221 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2281 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2341 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2401 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2461 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2521 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2581 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2641 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2701 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2761 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2821 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2881 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2941 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3001 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3061 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3121 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     3181 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3241 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3301 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     3361 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     3421 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     3481 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     3541 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     3601 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     3661 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     3721 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     3781 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     3841 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     3901 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     3961 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     4021 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     4081 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     4141 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4201 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4261 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4321 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4381 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4441 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4501 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     4561 cgcgcagccc gtactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     4621 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     4681 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     4741 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     4801 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     4861 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     4921 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     4981 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     5041 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     5101 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     5161 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5221 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5281 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     5341 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     5401 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     5461 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     5521 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     5581 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     5641 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     5701 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat
     5761 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat
     5821 accatgggcc atcatcatca tcatcacagc agcggcatga cttacaaact gatccttaac
     5881 ggtaaaactt tgaaaggcga aacca
//
