LOCUS       V045201                 7680 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045201
VERSION     V045201
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..2360
                     /label="Fbxl6(NM_013909)"
                     /note="Fbxl6(NM_013909)"
                     /gene="Fbxl6"
     CDS             2367..2432
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2456..3044
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3110..3158
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3360..3788
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3802..4131
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4198..4992
                     /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    5168..5289
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5338..5354)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5362..5378
                     /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(5386..5416)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5431..5452
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5740..6328)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6499..7359)
                     /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(7360..7464)
                     /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 tcccgtggcc tctaggcgag
      781 ttagacgtcg agttcgaagc tccaagcggc ccgatgcgcg gggccgctcg gcggaggact
      841 ggtggtggga ccgcctggca cctcgcggct ctgggtacca cctgctgcaa gcggacagca
      901 tgttgctcgt gctgccggac ctggagcctc cccgcgctcg tgcgcaccgg cgcgcccgtc
      961 gccgcgctcc gaggtcactg gctcgtggcc ccacggctgt cgccaagccc aggaccaaac
     1021 cccgaccgga gccgtcgctc gatcagggcc tggactctgg ttggggagac cgtattccct
     1081 tagaagtcct ggtgcatatt ttcgggttgt tggttgcggc tcatgggccc atgccgtttc
     1141 ttggcagggc tgcacgcgtt tgccgtcact ggcacgaagc cacctcccat ccttcactct
     1201 ggcatactgt gaccctgtca ccctcgctgg ttggccgggc tggcaagggc aaccttaagg
     1261 gagagaagaa gctccttgct tgcctggagt ggctcgtacc caatcggttc tctcagctcc
     1321 agagcttgac cctcatccac tggaagtctc aagtacactc tgtgttggag ctggttagca
     1381 agttctgccc tcggctcacc ttcctcaagc tttcagactg ccacactgtg actgctgaaa
     1441 ctctggtcat gttagcgaga gcctgctgcc agctccacag cctggaccta catcattcca
     1501 tggtggagtc cacagctgtg gtgagcttct tggaggaggc cgggtcccga atgcgtaaac
     1561 tgtggctgac ctacagttcc cagacgacgg ccatcttggg cgcactgctg gacaactgct
     1621 gcccacagct ccaagtcctc caggtcagca ctggcatgaa ctgcaacaac acacccctgc
     1681 agctgcctgt ggaagccctg cagaaaggct gcccccagct gcaggtgctg cggctgctga
     1741 atctgatttg gcttcccaag ccgtgtggcc gaggcgtgcc ccagggacca ggcttcccca
     1801 gtcttgagga gctctgtttg gctggctcca cctgcaactt tgtgagcaat gaggttctgg
     1861 gccgattgct ccatcgctcc ccgaaactgc gcatgctgga ccttcgaggc tgcgctaggg
     1921 tcactcctag cggtctgtgt catctgccat gtcaagagct ggagcaactc tacctgggcc
     1981 tgtatggcat atctgatggc ttgaccctgg ctaaggatgg cagccccctg ttgacccgga
     2041 agtggtatca caccctgagg gagctggact tcagtggcca aggcttcagc gagaaggact
     2101 tggaacaggc cctggctgtt ttctcgggca cccctggggg cttacaccca gccttgtgct
     2161 ccctcaacct gaggggtacc cgagttaccc caagcacggt cagttctgtg attagtggtt
     2221 gcccggggct gctgtatctt aacctggagt cctgtcgttg cctcccccga ggtctgaagc
     2281 gcgtctaccg gggcctggag gaagtccagt ggtgtctaga gcagctactt accagcccgc
     2341 cctctgccaa agagcccact ggatccgact acaaagacca tgacggtgat tataaagatc
     2401 atgacatcga ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca
     2461 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2521 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2581 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2641 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2701 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2761 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2821 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2881 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2941 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3001 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     3061 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3121 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3181 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     3241 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     3301 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     3361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3421 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3481 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     3541 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3661 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3721 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3781 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3841 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3961 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4021 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4081 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4141 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4201 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4261 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4321 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4381 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4441 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4501 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4561 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4621 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4681 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4741 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4801 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4861 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4921 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4981 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     5041 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5101 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5161 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5281 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5341 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5401 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5461 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5521 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5581 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5641 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5701 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5761 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5821 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5881 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5941 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6001 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6061 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6121 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6181 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6241 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6301 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6361 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7381 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7441 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7501 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     7561 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7621 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
