LOCUS       V032228                 8077 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032228
VERSION     V032228
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"
     CDS             683..2803
                     /label="FBXW7(NM_033632)"
                     /note="FBXW7(NM_033632)"
                     /gene="FBXW7"
     CDS             2810..2833
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    2853..3441
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3507..3555
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3757..4185
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4199..4528
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4595..5389
                     /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    5565..5686
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5735..5751)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5759..5775
                     /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(5783..5813)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5828..5849
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6137..6725)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6896..7756)
                     /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(7757..7861)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgaatca ggaactgctc tctgtgggca gcaaaagacg
      721 acgaactgga ggctctctga gaggtaaccc ttcctcaagc caggtagatg aagaacagat
      781 gaatcgtgtg gtagaggagg aacagcaaca gcaactcaga caacaagagg aggagcacac
      841 tgcaaggaat ggtgaagttg ttggagtaga acctagacct ggaggccaaa atgattccca
      901 gcaaggacag ttggaagaaa acaataatag atttatttcg gtagatgagg actcctcagg
      961 aaaccaagaa gaacaagagg aagatgaaga acatgctggt gaacaagatg aggaggatga
     1021 ggaggaggag gagatggacc aggagagtga cgattttgat cagtctgatg atagtagcag
     1081 agaagatgaa catacacata ctaacagtgt cacgaactcc agtagtattg tggacctgcc
     1141 cgttcaccaa ctctcctccc cattctatac aaaaacaaca aaaatgaaaa gaaagttgga
     1201 ccatggttct gaggtccgct ctttttcttt gggaaagaaa ccatgcaaag tctcagaata
     1261 tacaagtacc actgggcttg taccatgttc agcaacacca acaacttttg gggacctcag
     1321 agcagccaat ggccaagggc aacaacgacg ccgaattaca tctgtccagc cacctacagg
     1381 cctccaggaa tggctaaaaa tgtttcagag ctggagtgga ccagagaaat tgcttgcttt
     1441 agatgaactc attgatagtt gtgaaccaac acaagtaaaa catatgatgc aagtgataga
     1501 accccagttt caacgagact tcatttcatt gctccctaaa gagttggcac tctatgtgct
     1561 ttcattcctg gaacccaaag acctgctaca agcagctcag acatgtcgct actggagaat
     1621 tttggctgaa gacaaccttc tctggagaga gaaatgcaaa gaagagggga ttgatgaacc
     1681 attgcacatc aagagaagaa aagtaataaa accaggtttc atacacagtc catggaaaag
     1741 tgcatacatc agacagcaca gaattgatac taactggagg cgaggagaac tcaaatctcc
     1801 taaggtgctg aaaggacatg atgatcatgt gatcacatgc ttacagtttt gtggtaaccg
     1861 aatagttagt ggttctgatg acaacacttt aaaagtttgg tcagcagtca caggcaaatg
     1921 tctgagaaca ttagtgggac atacaggtgg agtatggtca tcacaaatga gagacaacat
     1981 catcattagt ggatctacag atcggacact caaagtgtgg aatgcagaga ctggagaatg
     2041 tatacacacc ttatatgggc atacttccac tgtgcgttgt atgcatcttc atgaaaaaag
     2101 agttgttagc ggttctcgag atgccactct tagggtttgg gatattgaga caggccagtg
     2161 tttacatgtt ttgatgggtc atgttgcagc agtccgctgt gttcaatatg atggcaggag
     2221 ggttgttagt ggagcatatg attttatggt aaaggtgtgg gatccagaga ctgaaacctg
     2281 tctacacacg ttgcaggggc atactaatag agtctattca ttacagtttg atggtatcca
     2341 tgtggtgagt ggatctcttg atacatcaat ccgtgtttgg gatgtggaga cagggaattg
     2401 cattcacacg ttaacagggc accagtcgtt aacaagtgga atggaactca aagacaatat
     2461 tcttgtctct gggaatgcag attctacagt taaaatctgg gatatcaaaa caggacagtg
     2521 tttacaaaca ttgcaaggtc ccaacaagca tcagagtgct gtgacctgtt tacagttcaa
     2581 caagaacttt gtaattacca gctcagatga tggaactgta aaactatggg acttgaaaac
     2641 gggtgaattt attcgaaacc tagtcacatt ggagagtggg gggagtgggg gagttgtgtg
     2701 gcggatcaga gcctcaaaca caaagctggt gtgtgcagtt gggagtcgga atgggactga
     2761 agaaaccaag ctgctggtgc tggactttga tgtggacatg aagctcgagc accatcacca
     2821 tcaccatcac cattagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg
     2881 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2941 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3001 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3061 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3121 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3181 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3241 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3301 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3361 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3421 ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac
     3481 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     3541 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     3601 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     3661 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     3721 cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3901 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3961 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4081 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct
     4141 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4201 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4261 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4321 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4381 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4441 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4501 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4561 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4621 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4681 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4741 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4801 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4861 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4921 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4981 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5041 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5101 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5161 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5221 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5281 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5341 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5401 gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5461 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5521 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5701 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5761 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5821 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5881 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5941 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6001 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6061 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6301 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     6361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6541 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6601 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6661 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6721 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6781 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7741 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7801 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7861 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     7921 atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     7981 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     8041 aggcttgacc gacaattgca tgaagaatct gcttagg
//
