LOCUS       V032229                 8086 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032229
VERSION     V032229
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..2812
                     /label="Fbxw7(NM_001177774)"
                     /note="Fbxw7(NM_001177774)"
                     /gene="Fbxw7"
     CDS             2819..2842
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    2862..3450
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3516..3564
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3766..4194
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4208..4537
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4604..5398
                     /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    5574..5695
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5744..5760)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5768..5784
                     /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(5792..5822)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5837..5858
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6146..6734)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6905..7765)
                     /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(7766..7870)
                     /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 gagggaatgc ttcctcaagc caggttgatg agggacagat
      781 gaatcgcgtg gttgaggagg atccacagca gcaagcgaga catcaagagg aggagcacac
      841 tgcgcggaat ggtgaacttg tgggtgcaaa ccctaggcct ggagaccaga acgataccca
      901 gcaaggacaa gtggaggaaa ataataaccg ctttatttca gtagatgagg actcttcggg
      961 aaatcaggaa gagcaagagg aagatgaaga gcatgctggg gaacaggagg aggaagagga
     1021 ggaagaggaa gaggaggagg agatggacca ggagagtgat gattttgatc cgtctgatga
     1081 cagtagcaga gaagatgaac atacgcacaa tagcaatgtc acaaactgca gtagtgtctc
     1141 ggacctgccc gctcaccagc tctcctctcc attctataca aagacaacaa aaatgaaaag
     1201 aaagttggac catggttctg aagttcgttc cttttctttg ggaaagaaac catgcaaagt
     1261 ctcagattat accagtacca ctggccttgt accatgttca gcaacaccaa caacttttgg
     1321 ggacctgaga gcagccaatg ggcaagggca gcagcggcgg aggattacat ctgtccaacc
     1381 acccacaggc cttcaagagt ggctgaaaat gtttcagagc tggagcggac cagagaagtt
     1441 gctggcttta gatgagctca ttgacagctg tgaaccaaca caagtgaagc atatgatgca
     1501 agtgatagag ccccagttcc agcgagactt catctccttg cttcctaaag agttggcact
     1561 ctatgtgctt tcattcctgg aacccaaaga cctgctgcaa gcggctcaga cttgtcgata
     1621 ctggagaatt ttggctgagg ataaccttct ctggagagag aaatgtaaag aagaggggat
     1681 tgatgaaccg ttgcacatca agagaagaaa aataataaaa ccaggtttca tacacagccc
     1741 atggaagagt gcgtatatca gacagcacag aattgataca aactggagac gaggagaact
     1801 caaatctcct aaggtgctga aagggcatga tgaccatgtg atcacatgcc tacagttttg
     1861 tggcaaccgc atagttagtg gttctgatga caacacttta aaagtttggt cagcggtcac
     1921 gggcaagtgt ctgagaacgt tagtgggaca tacaggtgga gtgtggtcat cacagatgag
     1981 agacaatatc atcatcagtg gatcgactga ccggactctc aaagtgtgga atgctgaaac
     2041 tggagagtgt atacatactt tatatgggca cacttctact gtacggtgta tgcatctcca
     2101 tgaaaaaagg gttgtaagcg gttctcgaga tgccactctc agggtttggg atattgagac
     2161 cggccagtgt ttacacgtcc tgatgggtca cgtagcagcg gtccgctgcg ttcagtatga
     2221 tggcaggagg gttgttagtg gagcttatga ttttatggtg aaggtgtggg atccagagac
     2281 tgagacctgt ctacacacgt tacagggaca cactaataga gtctattcat tacagtttga
     2341 tggcatccat gtggtgagtg gatctcttga tacatcaatc cgagtctggg atgtggagac
     2401 agggaattgt attcacacgc taacaggaca ccagtcatta acgagtggaa tggaactcaa
     2461 agacaatatt cttgtctctg ggaatgcaga ttctacagtt aagatctggg atatcaaaac
     2521 aggacagtgt ttacaaactt tgcaaggtcc cagcaagcat cagagcgctg tgacctgctt
     2581 acagttcaac aagaacttcg taattaccag ctcagacgac ggaacggtca aactctggga
     2641 cttgaaaacg ggtgaattta tccgaaacct cgtcacattg gagagtgggg ggagcggggg
     2701 agttgtgtgg cggatcaggg cctcaaacac aaagctggtg tgtgcagtcg ggagtcggaa
     2761 tggaactgag gaaaccaagc tcctggtgct ggactttgat gtggacatga aactcgagca
     2821 ccatcaccat caccatcacc attagggccc gatatctcga caatcaacct ctggattaca
     2881 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2941 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3001 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3061 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3121 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3181 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3241 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3301 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3361 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3421 gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg
     3481 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     3541 gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     3601 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     3661 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     3721 ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3781 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3841 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3901 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     3961 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4021 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4081 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     4141 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4201 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4261 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4321 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4381 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4441 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4501 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4561 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4621 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4681 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4741 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4801 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4861 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4921 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4981 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5041 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5101 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5161 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5221 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5281 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5341 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5401 gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5461 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5521 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5581 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5641 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5701 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5761 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5821 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5881 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5941 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6001 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6061 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     6361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7921 cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     7981 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8041 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
