LOCUS       sequence-189864-        6144 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 6144)
  AUTHORS   Huibregtse JM, Scheffner M, Howley PM
  TITLE     A cellular protein mediates association of p53 with the E6 
            oncoprotein of human papillomavirus types 16 or 18.
  JOURNAL   EMBO J. 1991 Dec . 10(13):4129-35.
  PUBMED    1661671
REFERENCE   2  (bases 1 to 6144)
  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: "EMBO J. 
            1991 Dec . 10(13):4129-35."
COMMENT     Sequence Label: p3113 GST-p53
FEATURES             Location/Qualifiers
     source          1..6144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..32
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        105..182
                     /gene="lacI"
                     /label=lacI promoter
     CDS             183..1265
                     /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     complement(202..221)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1314..1319,1320..1337,1338..1344)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1352..1368
                     /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     1376..1392
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1376..1392
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1388..1915
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1408..1425)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1408..1424)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1417..1439)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2017..2022,2023..2038,2039..2045)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2053..2069
                     /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)."
     CDS             2092..2745
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2703..2725
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2752..2769
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2791..3969
                     /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="EEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLS
                     PDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQG
                     SYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYK
                     QSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPP
                     EVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDR
                     RTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMF
                     RELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(4028..4050)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4195..4213)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4281..4385
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4386..4454,4455..5246)
                     /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     complement(4604..4623)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5417..6005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5906..5925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
       61 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc
      121 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
      181 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
      241 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
      301 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
      361 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
      421 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
      481 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
      541 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
      601 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
      661 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
      721 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
      781 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
      841 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
      901 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
      961 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     1021 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     1081 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     1141 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     1201 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     1261 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac
     1321 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     1381 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
     1441 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1501 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1561 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
     1621 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
     1681 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
     1741 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     1801 ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa
     1861 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
     1921 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
     1981 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta
     2041 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt catgtcccct
     2101 atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat
     2161 cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac
     2221 aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt
     2281 aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt
     2341 ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga
     2401 tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt
     2461 agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta
     2521 aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta
     2581 tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt
     2641 gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg
     2701 cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga tctggttccg
     2761 cgtggatctc gtcgtgcatc tgttggatcc gaggagccgc agtcagatcc tagcgtcgag
     2821 ccccctctga gtcaggaaac attttcagac ctatggaaac tacttcctga aaacaacgtt
     2881 ctgtccccct tgccgtccca agcaatggat gatttgatgc tgtccccgga cgatattgaa
     2941 caatggttca ctgaagaccc aggtccagat gaagctccca gaatgccaga ggctgctccc
     3001 cccgtggccc ctgcaccagc agctcctaca ccggcggccc ctgcaccagc cccctcctgg
     3061 cccctgtcat cttctgtccc ttcccagaaa acctaccagg gcagctacgg tttccgtctg
     3121 ggcttcttgc attctgggac agccaagtct gtgacttgca cgtactcccc tgccctcaac
     3181 aagatgtttt gccaactggc caagacctgc cctgtgcagc tgtgggttga ttccacaccc
     3241 ccgcccggca cccgcgtccg cgccatggcc atctacaagc agtcacagca catgacggag
     3301 gttgtgaggc gctgccccca ccatgagcgc tgctcagata gcgatggtct ggcccctcct
     3361 cagcatctta tccgagtgga aggaaatttg cgtgtggagt atttggatga cagaaacact
     3421 tttcgacata gtgtggtggt gccctatgag ccgcctgagg ttggctctga ctgtaccacc
     3481 atccactaca actacatgtg taacagttcc tgcatgggcg gcatgaaccg gaggcccatc
     3541 ctcaccatca tcacactgga agactccagt ggtaatctac tgggacggaa cagctttgag
     3601 gtgcgtgttt gtgcctgtcc tgggagagac cggcgcacag aggaagagaa tctccgcaag
     3661 aaaggggagc ctcaccacga gctgccccca gggagcacta agcgagcact gcccaacaac
     3721 accagctcct ctccccagcc aaagaagaaa ccactggatg gagaatattt cacccttcag
     3781 atccgtgggc gtgagcgctt cgagatgttc cgagagctga atgaggcctt ggaactcaag
     3841 gatgcccagg ctgggaagga gccagggggg agcagggctc actccagcca cctgaagtcc
     3901 aaaaagggtc agtctacctc ccgccataaa aaactcatgt tcaagacaga agggcctgac
     3961 tcagactgag aattcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg
     4021 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4081 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     4141 ccatgaccca gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg
     4201 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4261 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4321 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4381 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4441 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4501 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4561 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4621 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4681 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4741 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4801 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4861 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4921 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4981 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5041 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5101 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5161 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5221 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5281 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5341 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5401 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5461 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5521 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5581 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5641 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5701 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5761 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5821 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5881 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5941 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6001 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6061 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6121 gagctgatac cgctcgccgc agcc
//
