LOCUS       V028056                 6883 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028056
VERSION     V028056
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      901..910
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             964..2436
                     /label="MDM2(NM_001367990)"
                     /note="MDM2(NM_001367990)"
                     /gene="MDM2"
     polyA_signal    2483..2707
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2753..3181
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3195..3524
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3591..4385
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4559..4680
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4729..4745)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4753..4769
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4777..4807)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4822..4843
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5131..5716)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5887..6747)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6748..6852)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg attacaagga tgacgacgat aagactaagc ttatgtccag tgtggtggaa
      961 ttcatgtgca ataccaacat gtctgtacct actgatggtg ctgtaaccac ctcacagatt
     1021 ccagcttcgg aacaagagac cctggttaga ccaaagccat tgcttttgaa gttattaaag
     1081 tctgttggtg cacaaaaaga cacttatact atgaaagagg ttctttttta tcttggccag
     1141 tatattatga ctaaacgatt atatgatgag aagcaacaac atattgtata ttgttcaaat
     1201 gatcttctag gagatttgtt tggcgtgcca agcttctctg tgaaagagca caggaaaata
     1261 tataccatga tctacaggaa cttggtagta gtcaatcagc aggaatcatc ggactcaggt
     1321 acatctgtga gtgagaacag gtgtcacctt gaaggtggga gtgatcaaaa ggaccttgta
     1381 caagagcttc aggaagagaa accttcatct tcacatttgg tttctagacc atctacctca
     1441 tctagaagga gagcaattag tgagacagaa gaaaattcag atgaattatc tggtgaacga
     1501 caaagaaaac gccacaaatc tgatagtatt tccctttcct ttgatgaaag cctggctctg
     1561 tgtgtaataa gggagatatg ttgtgaaaga agcagtagca gtgaatctac agggacgcca
     1621 tcgaatccgg atcttgatgc tggtgtaagt gaacattcag gtgattggtt ggatcaggat
     1681 tcagtttcag atcagtttag tgtagaattt gaagttgaat ctctcgactc agaagattat
     1741 agccttagtg aagaaggaca agaactctca gatgaagatg atgaggtata tcaagttact
     1801 gtgtatcagg caggggagag tgatacagat tcatttgaag aagatcctga aatttcctta
     1861 gctgactatt ggaaatgcac ttcatgcaat gaaatgaatc ccccccttcc atcacattgc
     1921 aacagatgtt gggcccttcg tgagaattgg cttcctgaag ataaagggaa agataaaggg
     1981 gaaatctctg agaaagccaa actggaaaac tcaacacaag ctgaagaggg ctttgatgtt
     2041 cctgattgta aaaaaactat agtgaatgat tccagagagt catgtgttga ggaaaatgat
     2101 gataaaatta cacaagcttc acaatcacaa gaaagtgaag actattctca gccatcaact
     2161 tctagtagca ttatttatag cagccaagaa gatgtgaaag agtttgaaag ggaagaaacc
     2221 caagacaaag aagagagtgt ggaatctagt ttgcccctta atgccattga accttgtgtg
     2281 atttgtcaag gtcgacctaa aaatggttgc attgtccatg gcaaaacagg acatcttatg
     2341 gcctgcttta catgtgcaaa gaagctaaag aaaaggaata agccctgccc agtatgtaga
     2401 caaccaattc aaatgattgt gctaacttat ttcccctagc tcgagtctag agggcccgtt
     2461 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2521 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2581 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2641 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2701 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2761 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2821 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2881 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2941 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3001 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3061 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3121 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3181 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3241 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3301 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3361 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3421 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3481 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3541 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3601 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3661 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3721 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3781 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3841 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3901 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3961 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4021 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4081 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4141 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4201 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4261 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4321 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4381 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4441 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4501 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4561 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4621 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4681 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4741 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4801 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4861 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5641 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     5701 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5761 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5821 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5881 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5941 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6001 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6061 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6121 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6181 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6241 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6301 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6361 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6421 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6481 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6541 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6601 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6661 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6721 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6781 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6841 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
