LOCUS       V046209                 7473 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046209
VERSION     V046209
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..2153
                     /label="Fbxw17(NM_175401)"
                     /note="Fbxw17(NM_175401)"
                     /gene="Fbxw17"
     CDS             2160..2225
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2249..2837
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2903..2951
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3153..3581
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3595..3924
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3991..4785
                     /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    4961..5082
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5131..5147)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5155..5171
                     /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(5179..5209)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5224..5245
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5533..6121)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6292..7152)
                     /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(7153..7257)
                     /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 ccaccatggc gccgcagctt ggcccgcacg
      781 aactgcagca catcttctcc ttcctggagg cccgggacct gctgcgcgct gcgcaggtga
      841 acaaggtctg gaatgaggtt tcgatgacca aggaactgtg gaggcagctg tgtctgaggc
      901 gatgggcctc ctgcaaggcc ttcccagtgg tcgtgggcac gcaaacatgg aagaagtact
      961 atttctgccg gtctgagcta gagttccgta tggaatctgg gcggccacag gatttcatct
     1021 gcaaagctat ttctgggcac acaggaatga tagatcagct ggcctacgtc tcaccccatg
     1081 agtaccgctt tgatgaaggg gcaaggtccg tggtttgcac tgtgtcttca gactgtactg
     1141 tgcgtgcctg ggatctgcag gagggctcag agatctggtc aagtgctgtg cagcctgctc
     1201 ctctggtgaa tgtggtggcc tatcctcagc tgcagctggt tgtcactgtg gacatacagg
     1261 gcttgatcat ggggtggaaa gcaaagaccg ggtctgagtg ggccttcttc agcttgccgg
     1321 catcctgttc ttcaatggag gcctgtgacc acccagaggg ccccttcttg ctggtagcct
     1381 gtgatgaagg gaccctctac actatgacag tacctcagct gcaagtggtc tccagagtac
     1441 ccacatttcc tcactcctct gtgagcctca cgtgctcccc tgacacacag tgggtatttg
     1501 tgtctgcaca aggctcagac ttaggcccaa aggtgttcta cactcagccc ttgctgcacc
     1561 catcagaaga cagacttcct gtctccacta cccttcccat caggctgagc tccagagcct
     1621 gctgggctcc ggatgaggca gccagactga tggtgataca cagagatgac ggtggcgtgc
     1681 atctggttat cactaccttc gagttaaagg ccaccaagtc cagggacaga ttgagcatcc
     1741 ttgcacagca gctggccagc ttccacctgc cggacactat gactcctcac ctcatgcagg
     1801 gccatggctc ccaaacgatc ctgctcacct ccaggtcaga gctggtgctc ttcaccatac
     1861 atggcctgca gctgatggcc ttccaggacc accagaggcc aatcacgtct atgtgggtgg
     1921 atccagaccg tgtcatcact gctgccttgg acctgtcctt gcgtgtctat gtgtggaata
     1981 aaaagaacac atcacctgtc ctcaagagct gttaccattt gctcggggga tctcacagat
     2041 gggccagtgg attcacccac gtgaagagtg acagcagaag cattgctggg gtggaagcca
     2101 gaagcactgg gacaagtatt ctgaggtcat attgcttcaa ggttaagctt gacggatccg
     2161 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     2221 acaagtagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag
     2281 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2341 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2401 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2461 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2521 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2581 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2641 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2701 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2761 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2821 ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata
     2881 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     2941 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3001 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3061 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     3121 tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3181 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3241 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3301 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3361 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3421 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3481 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3541 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3601 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3661 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3721 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3781 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3841 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3901 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3961 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4021 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4081 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4141 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4201 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4261 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4321 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4381 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4441 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4501 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4561 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4621 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4681 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4741 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4801 tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4861 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4921 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4981 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5041 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5101 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5161 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5221 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5281 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5341 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5401 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5461 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5521 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5581 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5641 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5701 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5761 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5821 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5881 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5941 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6001 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6061 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6121 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6181 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6241 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6301 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6361 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6421 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6481 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6541 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6601 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6661 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6721 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6781 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6841 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6901 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6961 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7021 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7081 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7141 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7201 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7261 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7321 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7381 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7441 ttgaccgaca attgcatgaa gaatctgctt agg
//
