LOCUS       sequence-327631-        6184 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 6184)
  AUTHORS   Ayed A, Mulder FA, Yi GS, Lu Y, Kay LE, Arrowsmith CH
  TITLE     Latent and active p53 are identical in conformation.
  JOURNAL   Nat Struct Biol. 2001 Sep . 8(9):756-60.
  PUBMED    11524676
REFERENCE   2  (bases 1 to 6184)
  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: "Nat Struct 
            Biol. 2001 Sep . 8(9):756-60."
COMMENT     Sequence Label: pGEX-human p53-(1-393)
FEATURES             Location/Qualifiers
     source          1..6184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(48..53,54..69,70..76)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    84..100
                     /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             123..776
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     734..756
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             783..800
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             819..2000
                     /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
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     primer_bind     complement(2099..2121)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2266..2284)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2352..2456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2457..2525,2526..3317)
                     /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(2675..2694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3488..4076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3977..3996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4230..4247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4320..4397
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4398..5480
                     /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(4417..4436)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5529..5534,5535..5552,5553..5559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5567..5583
                     /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     5591..5607
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5591..5607
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5603..6130
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5623..5640)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5623..5639)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5632..5654)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc
       61 atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagtat
      121 tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc
      181 ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata
      241 aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg
      301 atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca
      361 acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt
      421 tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag
      481 ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata
      541 aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg
      601 atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta
      661 aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag
      721 catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg
      781 atctggttcc gcgtggatct cgtcgtgcat ctgttcatat ggaggagccg cagtcagatc
      841 ctagcgtcga gccccctctg agtcaggaaa cattttcaga cctatggaaa ctacttcctg
      901 aaaacaacgt tctgtccccc ttgccgtccc aagcaatgga tgatttgatg ctgtccccgg
      961 acgatattga acaatggttc accgaagacc caggtccaga tgaagctccc agaatgccag
     1021 aggctgctcc ccccgtggcc cctgcaccag cagctcctac accggcggcc cctgcaccag
     1081 ccccctcctg gcccctgtca tcttctgtcc cttcccagaa aacctaccag ggcagctacg
     1141 gtttccgtct gggcttcttg cattctggga cagccaagtc tgtgacttgc acgtactccc
     1201 ctgccctcaa caagatgttt tgccaactgg ccaagacctg ccctgtgcag ctgtgggttg
     1261 attccacacc cccgcccggc acccgcgtcc gcgccatggc catctacaag cagtcacagc
     1321 acatgacgga ggttgtgagg cgctgccccc accatgagcg ctgctcagat agcgatggtc
     1381 tggcccctcc tcagcatctt atccgagtgg aaggaaattt gcgtgtggag tatttggatg
     1441 acagaaacac ttttcgacat agtgtggtgg tgccctatga gccgcctgag gttggctctg
     1501 actgtaccac catccactac aactacatgt gtaacagttc ctgcatgggc ggcatgaacc
     1561 ggaggcccat cctcaccatc atcacactgg aagactccag tggtaatcta ctgggacgga
     1621 acagctttga ggtgcgtgtt tgtgcctgtc ctgggagaga ccggcgcaca gaggaagaga
     1681 atctccgcaa gaaaggggag cctcaccacg agctgccccc agggagcact aagcgagcac
     1741 tgcccaacaa caccagctcc tctccccagc caaagaagaa accactggat ggagaatatt
     1801 tcacccttca gatccgtggg cgtgagcgct tcgagatgtt ccgagagctg aatgaggcct
     1861 tggaactcaa ggatgcccag gctgggaagg agccaggggg gagcagggct cactccagcc
     1921 acctgaagtc caaaaagggt cagtctacct cccgccataa aaaactcatg ttcaagacag
     1981 aagggcctga ctcagactga cattctccac ttcttgttcc ccactgacag ggatccccgg
     2041 gaattcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     2101 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     2161 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     2221 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
     2281 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2341 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2401 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2461 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2521 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2581 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2641 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     2701 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2761 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2821 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2881 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2941 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3001 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3061 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3121 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3181 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3241 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3301 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3361 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3421 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3481 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3601 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4081 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4141 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4201 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4261 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
     4321 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     4381 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     4441 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     4501 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     4561 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     4621 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     4681 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     4741 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     4801 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     4861 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     4921 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     4981 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     5041 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     5101 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     5161 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     5221 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     5281 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     5341 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     5401 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5461 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     5521 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5581 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
     5641 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5701 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5761 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
     5821 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
     5881 gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc
     5941 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     6001 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac
     6061 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg
     6121 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
     6181 tttt
//
