LOCUS       sequence-378454-        5299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human MDM2 (17-125) in e. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5299)
  AUTHORS   Borcherds W, Theillet FX, Katzer A, Finzel A, Mishall KM, Powell AT,
            Wu H, Manieri W, Dieterich C, Selenko P, Loewer A, Daughdrill GW
  TITLE     Disorder and residual helicity alter p53-Mdm2 binding affinity and 
            signaling in cells.
  JOURNAL   Nat Chem Biol. 2014 Dec;10(12):1000-2. doi: 10.1038/nchembio.1668. 
            Epub 2014 Nov 2.
  PUBMED    25362358
REFERENCE   2  (bases 1 to 5299)
  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: 
            "10.1038/nchembio.1668"; journalName: "Nat Chem Biol"; date: 
            "2014-12"; volume: "10"; issue: "12"; pages: "1000-2"
COMMENT     Sequence Label: pGEX6P2-MDM2 (17-125)
FEATURES             Location/Qualifiers
     source          1..5299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..46
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        119..196
                     /gene="lacI"
                     /label=lacI promoter
     CDS             197..1279
                     /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(216..235)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1328..1333,1334..1351,1352..1358)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1366..1382
                     /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     1390..1406
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1390..1406
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1402..1929
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1422..1439)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1422..1438)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1431..1453)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2031..2036,2037..2052,2053..2059)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2067..2083
                     /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             2106..2759
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2717..2739
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2766..2789
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3197..3219)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3364..3382)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3450..3554
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3555..3623,3624..4415)
                     /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(3773..3792)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4586..5174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5075..5094
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
       61 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
      121 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
      181 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
      241 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
      301 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
      361 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
      421 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
      481 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
      541 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
      601 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
      661 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
      721 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
      781 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
      841 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
      901 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
      961 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     1021 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     1081 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     1141 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc acgctagcgc ccaatacgca
     1201 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     1261 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     1321 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     1381 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
     1441 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1501 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1561 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
     1621 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
     1681 cacggttacg atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg
     1741 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
     1801 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaattag cttatcgact
     1861 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
     1921 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     1981 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     2041 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2101 tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac
     2161 ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg
     2221 ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata
     2281 ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc
     2341 acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg
     2401 ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca
     2461 aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc
     2521 ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc
     2581 ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt
     2641 ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata
     2701 tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat
     2761 cggatctgga agttctgttc caggggcccc tgggatccag ccagattccg gcaagcgaac
     2821 aagaaaccct ggttcgtccg aaaccgctgc tgctgaaact gctgaaaagc gttggtgcac
     2881 agaaagatac ctataccatg aaagaggtgc tgttttatct gggccagtat attatgacca
     2941 aacgcctgta tgatgagaaa cagcagcata ttgtgtattg cagcaatgat ctgctgggtg
     3001 acctgtttgg tgttccgagc tttagcgtta aagaacaccg taaaatctat accatgatct
     3061 atcgtaatct ggtggtggtt aatcagcaag aaagcagcga tagcggcacc agcgttagcg
     3121 aaaactaata actcgagcgg ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg
     3181 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3241 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3301 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa
     3361 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3421 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3481 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3541 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3601 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3661 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3721 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3781 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3841 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3901 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3961 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4021 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4081 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     4141 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4201 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4261 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4321 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4381 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4441 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4501 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4561 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4621 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4681 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4741 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4801 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4861 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4921 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4981 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5041 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5101 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5161 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5221 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5281 cctttgagtg agctgatac
//
