LOCUS       V037211                 7566 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037211
VERSION     V037211
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2246
                     /label="MDM2(NM_002392)"
                     /note="MDM2(NM_002392)"
                     /gene="MDM2"
     CDS             2253..2318
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2342..2930
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2996..3044
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3246..3674
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3688..4017
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4084..4878
                     /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    5054..5175
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5224..5240)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5248..5264
                     /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(5272..5302)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5317..5338
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5626..6214)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6385..7245)
                     /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(7246..7350)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt gaggagcagg caaatgtgca
      781 ataccaacat gtctgtacct actgatggtg ctgtaaccac ctcacagatt ccagcttcgg
      841 aacaagagac cctggttaga ccaaagccat tgcttttgaa gttattaaag tctgttggtg
      901 cacaaaaaga cacttatact atgaaagagg ttctttttta tcttggccag tatattatga
      961 ctaaacgatt atatgatgag aagcaacaac atattgtata ttgttcaaat gatcttctag
     1021 gagatttgtt tggcgtgcca agcttctctg tgaaagagca caggaaaata tataccatga
     1081 tctacaggaa cttggtagta gtcaatcagc aggaatcatc ggactcaggt acatctgtga
     1141 gtgagaacag gtgtcacctt gaaggtggga gtgatcaaaa ggaccttgta caagagcttc
     1201 aggaagagaa accttcatct tcacatttgg tttctagacc atctacctca tctagaagga
     1261 gagcaattag tgagacagaa gaaaattcag atgaattatc tggtgaacga caaagaaaac
     1321 gccacaaatc tgatagtatt tccctttcct ttgatgaaag cctggctctg tgtgtaataa
     1381 gggagatatg ttgtgaaaga agcagtagca gtgaatctac agggacgcca tcgaatccgg
     1441 atcttgatgc tggtgtaagt gaacattcag gtgattggtt ggatcaggat tcagtttcag
     1501 atcagtttag tgtagaattt gaagttgaat ctctcgactc agaagattat agccttagtg
     1561 aagaaggaca agaactctca gatgaagatg atgaggtata tcaagttact gtgtatcagg
     1621 caggggagag tgatacagat tcatttgaag aagatcctga aatttcctta gctgactatt
     1681 ggaaatgcac ttcatgcaat gaaatgaatc ccccccttcc atcacattgc aacagatgtt
     1741 gggcccttcg tgagaattgg cttcctgaag ataaagggaa agataaaggg gaaatctctg
     1801 agaaagccaa actggaaaac tcaacacaag ctgaagaggg ctttgatgtt cctgattgta
     1861 aaaaaactat agtgaatgat tccagagagt catgtgttga ggaaaatgat gataaaatta
     1921 cacaagcttc acaatcacaa gaaagtgaag actattctca gccatcaact tctagtagca
     1981 ttatttatag cagccaagaa gatgtgaaag agtttgaaag ggaagaaacc caagacaaag
     2041 aagagagtgt ggaatctagt ttgcccctta atgccattga accttgtgtg atttgtcaag
     2101 gtcgacctaa aaatggttgc attgtccatg gcaaaacagg acatcttatg gcctgcttta
     2161 catgtgcaaa gaagctaaag aaaaggaata agccctgccc agtatgtaga caaccaattc
     2221 aaatgattgt gctaacttat ttccccggat ccgactacaa agaccatgac ggtgattata
     2281 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     2341 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2401 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2461 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2521 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2581 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2641 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2701 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     2761 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2821 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2881 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     2941 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3001 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3061 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3121 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3181 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3241 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3301 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3361 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3421 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3481 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3541 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3601 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3661 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3781 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3841 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3901 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3961 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4021 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4081 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4141 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4201 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4261 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4321 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4381 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4441 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4501 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4561 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4621 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4681 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4741 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4801 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4861 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     4921 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4981 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5041 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5101 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5161 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5221 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5281 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5341 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5401 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5461 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5521 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5581 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5641 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5701 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5761 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5821 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5881 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5941 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6001 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6061 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6121 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6181 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6241 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6301 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6361 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6421 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6481 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6541 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6601 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6661 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6721 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6781 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6841 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6901 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6961 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7021 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7081 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7141 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7201 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7261 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7321 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7381 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     7441 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7501 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7561 cttagg
//
