LOCUS       sequence-129513-        6448 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 6448)
  AUTHORS   Zhou BP, Liao Y, Xia W, Zou Y, Spohn B, Hung MC
  TITLE     HER-2/neu induces p53 ubiquitination via Akt-mediated MDM2 
            phosphorylation.
  JOURNAL   Nat Cell Biol. 2001 Nov . 3(11):973-82.
  PUBMED    11715018
REFERENCE   2  (bases 1 to 6448)
  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 Cell 
            Biol. 2001 Nov . 3(11):973-82."
COMMENT     Sequence Label: pGEX-4T MDM2 WT
FEATURES             Location/Qualifiers
     source          1..6448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..88
                     /gene="lacI"
                     /label=lacI promoter
     CDS             89..1171
                     /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(108..127)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1220..1225,1226..1243,1244..1250)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1258..1274
                     /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     1282..1298
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1282..1298
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1294..1821
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1314..1331)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1314..1330)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1323..1345)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1923..1928,1929..1944,1945..1951)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1959..1975
                     /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             1998..2651
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2609..2631
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2658..2675
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4238..4260)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4405..4423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4491..4595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4596..4664,4665..5456)
                     /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(4814..4833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5627..6215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6116..6135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6369..6386
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cataaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
       61 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
      121 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
      181 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
      241 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
      301 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
      361 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
      421 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
      481 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
      541 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
      601 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
      661 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
      721 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
      781 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
      841 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
      901 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
      961 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     1021 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     1081 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1141 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
     1201 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     1261 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc
     1321 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1381 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1441 caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg
     1501 gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca
     1561 aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg
     1621 gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat
     1681 gttgatgaaa gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt
     1741 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
     1801 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
     1861 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
     1921 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
     1981 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
     2041 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
     2101 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
     2161 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
     2221 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
     2281 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
     2341 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
     2401 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
     2461 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
     2521 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
     2581 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
     2641 atcctccaaa atcggatctg gttccgcgtg gatccgccac catgtgcaat accaacatgt
     2701 ctgtacctac tgatggtgct gtaaccacct cacagattcc agcttcggaa caagagaccc
     2761 tggttagacc aaagccattg cttttgaagt tattaaagtc tgttggtgca caaaaagaca
     2821 cttatactat gaaagaggtt cttttttatc ttggccagta tattatgact aaacgattat
     2881 atgatgagaa gcaacaacat attgtatatt gttcaaatga tcttctagga gatttgtttg
     2941 gcgtgccaag cttctctgtg aaagagcaca ggaaaatata taccatgatc tacaggaact
     3001 tggtagtagt caatcagcag gaatcatcgg actcaggtac atctgtgagt gagaacaggt
     3061 gtcaccttga aggtgggagt gatcaaaagg accttgtaca agagcttcag gaagagaaac
     3121 cttcatcttc acatttggtt tctagaccat ctacctcatc tagaaggaga gcaattagtg
     3181 agacagaaga aaattcagat gaattatctg gtgaacgaca aagaaaacgc cacaaatctg
     3241 atagtatttc cctttccttt gatgaaagcc tggctctgtg tgtaataagg gagatatgtt
     3301 gtgaaagaag cagtagcagt gaatctacag ggacgccatc gaatccggat cttgatgctg
     3361 gtgtaagtga acattcaggt gattggttgg atcaggattc agtttcagat cagtttagtg
     3421 tagaatttga agttgaatct ctcgactcag aagattatag ccttagtgaa gaaggacaag
     3481 aactctcaga tgaagatgat gaggtatatc aagttactgt gtatcaggca ggggagagtg
     3541 atacagattc atttgaagaa gatcctgaaa tttccttagc tgactattgg aaatgcactt
     3601 catgcaatga aatgaatccc ccccttccat cacattgcaa cagatgttgg gcccttcgtg
     3661 agaattggct tcctgaagat aaagggaaag ataaagggga aatctctgag aaagccaaac
     3721 tggaaaactc aacacaagct gaagagggct ttgatgttcc tgattgtaaa aaaactatag
     3781 tgaatgattc cagagagtca tgtgttgagg aaaatgatga taaaattaca caagcttcac
     3841 aatcacaaga aagtgaagac tattctcagc catcaacttc tagtagcatt atttatagca
     3901 gccaagaaga tgtgaaagag tttgaaaggg aagaaaccca agacaaagaa gagagtgtgg
     3961 aatctagttt gccccttaat gccattgaac cttgtgtgat ttgtcaaggt cgacctaaaa
     4021 atggttgcat tgtccatggc aaaacaggac atcttatggc ctgctttaca tgtgcaaaga
     4081 agctaaagaa aaggaataag ccctgcccag tatgtagaca accaattcaa atgattgtgc
     4141 taacttattt cccctaggaa ttcccgggtc gactcgagcg gccgcatcgt gactgactga
     4201 cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4261 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4321 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     4381 tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4441 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4501 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4561 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4621 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4681 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4741 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4801 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     4861 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4921 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4981 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5041 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5101 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     5161 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5221 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5281 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5341 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5401 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5461 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5521 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5761 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     6421 ttacgcatct gtgcggtatt tcacaccg
//
