LOCUS       sequence-456540-        8427 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 8427)
  AUTHORS   Oliner JD, Kinzler KW, Meltzer PS, George DL, Vogelstein B
  TITLE     Amplification of a gene encoding a p53-associated protein in human 
            sarcomas.
  JOURNAL   Nature. 1992 Jul 2. 358(6381):80-3.
  PUBMED    1614537
REFERENCE   2  (bases 1 to 8427)
  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: "Nature. 
            1992 Jul 2. 358(6381):80-3."
COMMENT     Sequence Label: pCMV MDM-2
FEATURES             Location/Qualifiers
     source          1..8427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(315..339)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(336..539)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(365..385)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        540..919
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1212..1259
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1284..1303
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1497..2291)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1608..1627)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2218..2237
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2348..2493)
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(2904..2921)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2904..2920)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2913..2935)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3129..3148)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3248..3270
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3308..3326)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3394..3498
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3499..3567,3568..4359)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3717..3736)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4530..5118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5019..5038
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5272..5289
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5406..5427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5442..5447,5448..5465,5466..5472)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5480..5496
                     /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     5485..5507
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5504..5520
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5504..5520
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5860..5879
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    5878..5933
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     5979..5998
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          join(8079..8427,1..224)
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 ctaaagttta acgaaaaagt tacagaaaat gaaaagaaaa taagaggaga caatggttgt
       61 caacagagta gaaagtgaaa gaaacaaaat tatcatgagg gtccatggtg atacaaggga
      121 catcttccca ttctaaacaa caccctgaaa actttgcccc ctccatataa catgaatttt
      181 acaatagcga aaaagaaaga acaatcaagg gtccccaaac tcaccctgaa gttctcagga
      241 tcggtcgacc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
      301 tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata gacctcccac
      361 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
      421 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
      481 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
      541 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
      601 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
      661 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
      721 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
      781 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
      841 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
      901 ctagtcaata atcaatgtca acatggcggt aatgttggac atgagccaat ataaatgtac
      961 atattatgat atggatacaa cgtatgcaat gggccaagct ctctagaggg ccgcgatccg
     1021 ccgtccctaa cccacggggc ccgtggctat ggcagggctt gccgccccga cgttggctgc
     1081 gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc
     1141 gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     1201 cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt
     1261 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tcccccatct
     1321 cccggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     1381 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     1441 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     1501 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     1561 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     1621 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     1681 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     1741 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     1801 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     1861 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     1921 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     1981 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     2041 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     2101 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     2161 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     2221 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     2281 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatctgcg gcacgctgtt
     2341 gacgctgtta agcgggtcgc tgcagggtcg ctcggtgttc gaggccacac gcgtcacctt
     2401 aatatgcgaa gtggacctgg gaccgcgccg ccccgactgc atctgcgtgt tcgaattcgc
     2461 caatgacaag acgctgggcg gggtttgtgt catcatagaa ctaaagacat gcaaatatat
     2521 ttcttccggg gacaccgcca gcaaacgcga gcaacgggcc acggggatga agcaggatct
     2581 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
     2641 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
     2701 aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc
     2761 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     2821 tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg
     2881 cttgcggcag cgtgaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2941 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3001 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3061 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     3121 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3181 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3241 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3301 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3361 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3421 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3481 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3541 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3601 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3661 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3721 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3781 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3841 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3901 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3961 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4021 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4081 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4141 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4201 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4261 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4321 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4381 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4441 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4501 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4561 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4621 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4681 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4741 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4801 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4861 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4921 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4981 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5041 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5101 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5161 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5221 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5281 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5341 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5401 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     5461 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     5521 atgattacga attgcggccg atccgtcgag ggatctccat aagagaagag ggacagctat
     5581 gactgggagt agtcaggaga ggaggaaaaa tctggctagt aaaacatgta aggaaaattt
     5641 tagggatgtt aaagaaaaaa ataacacaaa acaaaatata aaaaaaatct aacctcaagt
     5701 caaggctttt ctatggaata aggaatggac agcagggggc tgtttcatat actgatgacc
     5761 tctttatagc caacctttgt tcatggcagc cagcatatgg gcatatgttg ccaaactcta
     5821 aaccaaatac tcattctgat gttttaaatg atttgccctc ccatatgtcc ttccgagtga
     5881 gagacacaaa aaattccaac acactattgc aatgaaaata aatttccttt attagccaga
     5941 agtcagatgc tcaaggggct tcatgatgtc cccataattt ttggcagagg gaaaaagatc
     6001 tcagtggtat ttgtgagcca gggcattggc cacaccagcc accaccttct gataggcagc
     6061 ctgcacctga ggagtgaatt cctcgacgga tcccccggga ggcagagctt gcagtgagcc
     6121 aagatcacgc cactgcactc cagcctgggt aacagagcaa gactcggtct caaaaaaaat
     6181 aataagttac atttaaatgt catatacata tttaagaaaa aaaaaaacca agtacttctc
     6241 atttaagaca gagtagaaat tatttaaaat taggagttgg tgtaaaggat gagctacata
     6301 ttcaagtcaa attatagtaa gtattcacta ttccactacc aaagtaggtc aattatacta
     6361 aagagaagaa atctatgtga attgaggcat tttctcactt tgatatatgt gaataaattt
     6421 caggttgtct aaattcctag ggttatatag ttagaaatat ataattctct tatagacagg
     6481 tcaactaggg gaaataagtt agcacaatca tttgaattgg ttgtctacat actgggcagg
     6541 gcttattcct tttctttagc ttctttgcac atgtaaagca ggccataaga tgtcctgttt
     6601 tgccatggac aatgcaacca tttttaggtc gaccttgaca aatcacacaa ggttcaatgg
     6661 cattaagggg caaactagat tccacactct cttctttgtc ttgggtttct tccctttcaa
     6721 actctttcac atcttcttgg ctgctataaa taatgctact agaagttgat ggctgagaat
     6781 agtcttcact ttcttgtgat tgtgaagctt gtgtaatttt atcatcattt tcctcaacac
     6841 atgactctct ggaatcattc actatagttt ttttacaatc aggaacatca aagccctctt
     6901 cagcttgtgt tgagttttcc agtttggctt tctcagagat ttccccttta tctttccctt
     6961 tatcttcagg aagccaattc tcacgaaggg cccaacatct gttgcaatgt gatggaaggg
     7021 ggggattcat ttcattgcat gaagtgcatt tccaatagtc agctaaggaa atttcaggat
     7081 cttcttcaaa tgaatctgta tcactctccc ctgcctgata cacagtaact tgatatacct
     7141 catcatcttc atctgagagt tcttgtcctt cttcactaag gctataatct tctgagtcga
     7201 gagattcaac ttcaaattct acactaaact gatctgaaac tgaatcctga tccaaccaat
     7261 cacctgaatg ttcacttaca ccagcatcaa gatccggatt cgatggcgtc cctgtagatt
     7321 cactgctact gcttctttca caacatatct cccttattac acacagagcc aggctttcat
     7381 caaaggaaag ggaaatacta tcagatttgt ggcgttttct ttgtcgttca ccagataatt
     7441 catctgaatt ttcttctgtc tcactaattg ctctccttct agatgaggta gatggtctag
     7501 aaaccaaatg tgaagatgaa ggtttctctt cctgaagctc ttgtacaagg tccttttgat
     7561 cactcccacc ttcaaggtga cacctgttct cactcacaga tgtacctgag tccgatgatt
     7621 cctgctgatt gactactacc aagttcctgt agatcatggt atatattttc ctgtgctctt
     7681 tcacagagaa gcttggcacg ccaaacaaat ctcctagaag atcatttgaa caatatacaa
     7741 tatgttgttg cttctcatca tataatcgtt tagtcataat atactggcca agataaaaaa
     7801 gaacctcttt catagtataa gtgtcttttt gtgcaccaac agactttaat aacttcaaaa
     7861 gcaatggctt tggtctaacc agggtctctt gttccgaagc tggaatctgt gaggtggtta
     7921 cagcaccatc agtaggtaca gacatgttgg tattgcacat ttgtttttag gttcgctttt
     7981 ctgttttatt tgtttcgaaa actgtcgacc gcggatccgt cgaggaattc tttgccaaaa
     8041 tgatgagaca gcacaacaac cagcacgttg cccaggagct gtaggaaaga gaagaaggca
     8101 tgaacatggt tagcagaggg gcccggtttg gactcagagt attttatcct catctcaaac
     8161 agtgtatatc attgtaacca taaagagaaa ggcaggatga tgaccagggt gtagttgttt
     8221 ctaccaataa gaatatttcc acgccagcca gaatttatat gcagaaatat tctaccttat
     8281 catttaatta taacaattgt tctctaaaac tgtgctgaag tacaatataa tataccctga
     8341 ttgccttgaa aaaaaagtga ttagagaaag tacttacaat ctgacaaata aacaaaagtg
     8401 aatttaaaaa ttcgttacaa atgcaag
//
