LOCUS       sequence-393114-        5962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of P53 TETCTD for segmental labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5962)
  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 5962)
  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: H6GB1tev-IntC-P53(304-393)
FEATURES             Location/Qualifiers
     source          1..5962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..99
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     183..202
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        183..201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    202..226
                     /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             241..263
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             276..293
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             309..470
                     /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             495..515
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             876..893
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(946..964)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      960..1007
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1044..1499
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1131..1150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1341..1362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1525..1629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1630..1698,1699..2490)
                     /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(1848..1867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2661..3249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3150..3169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3403..3420
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3435..3574
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3500..3519)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3660..3682
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3676..3867)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4676..5758)
                     /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     5720..5739
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5759..5836)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
       61 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
      121 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa
      181 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
      241 tttgtttaac tttaagaagg agatatacca tgggccatca tcatcatcat cacagcagcg
      301 gcatgactta caaactgatc cttaacggta aaactttgaa aggcgaaacc actaccgaag
      361 ctgttgatgc tgcgactgca gaaaaagttt tcaaacagta cgctaacgac aacggtgttg
      421 acggtgaatg gacttacgac gatgcgacta agaccttcac cgttactgaa gaattcagca
      481 gcggcagcag cggcgaaaac ctgtattttc agggctccca tatggaccat aattttgcac
      541 tcaaaaatgg cttcatagct tctaattgtt tcaatggtac taagcgagca ctgcccaaca
      601 acaccagctc ctctccccag ccaaagaaga aaccactgga tggagaatat ttcacccttc
      661 agatccgtgg gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc ttggaactca
      721 aggatgccca ggctgggaag gagccagggg ggagcagggc tcactccagc cacctgaagt
      781 ccaaaaaggg tcagtctacc tcccgccata aaaaactcat gttcaagaca gaagggcctg
      841 actcagacta atagaagctt gcggccgcac tcgagcacca ccaccaccac cactgagatc
      901 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
      961 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     1021 ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1081 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1141 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1201 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1261 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1321 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     1381 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     1441 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     1501 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1561 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1621 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1681 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1741 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1801 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1861 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1921 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1981 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2041 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2101 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2161 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2221 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2281 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2341 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2401 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2461 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2521 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2581 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3361 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3421 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3481 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3541 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3601 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3661 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3721 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3781 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3841 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     3901 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3961 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     4021 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     4081 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     4141 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     4201 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     4261 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     4321 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     4381 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     4441 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     4501 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     4561 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     4621 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     4681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4741 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     4801 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     4861 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     4921 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgt
     4981 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     5041 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     5101 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     5161 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     5221 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     5281 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     5341 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     5401 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     5461 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     5521 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     5581 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     5641 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     5701 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     5761 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     5821 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     5881 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     5941 gagatggcgc ccaacagtcc cc
//
