LOCUS       V018821                 7132 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018821
VERSION     V018821
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             991..3294
                     /label="Cul3(NM_016716)"
                     /note="Cul3(NM_016716)"
                     /gene="Cul3"
     CDS             3298..3327
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3346..3369
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3389..3405)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3416..4038
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4187..4775)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5104..5151
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5383..6177)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(6212..6569)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6571..6675)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6702..7130
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgccacc atgtcgaatc tgagcaaagg cacgggcagc
     1021 cggaaggaca ccaagatgcg gatccgggcc tttccgatga ccatggatga aaaatatgta
     1081 aacagcattt gggaccttct gaaaaatgca attcaagaaa tccagcgtaa gaataacagt
     1141 ggtcttagtt ttgaggagct ctatagaaat gcatatacaa tggttttgca taaacatgga
     1201 gaaaagctct acactggact aagagaagtt gttactgaac atctcataaa taaggtgcga
     1261 gaagatgtac taaattcatt gaataacaac tttcttcaaa cactaaatca agcttggaat
     1321 gatcatcaaa cagccatggt gatgattaga gacatattaa tgtatatgga ccgtgtatat
     1381 gtacaacaga ataatgtaga aaatgtctac aatttgggat taataatttt tcgagatcaa
     1441 gttgtacggt atgggtgtat tagggatcat cttcggcaaa ctttattgga tatgattgcc
     1501 agagagcgga aaggagaagt tgtagacaga ggagcaataa gaaatgcttg ccagatgtta
     1561 atgattttag gtcttgaagg aagatcagtc tatgaggaag attttgaggc tccgtttttg
     1621 gaaatgtctg cagaattttt tcagatggaa agccagaaat ttttagcaga aaacagtgct
     1681 tcagtatata taaagaaagt agaagctagg attaatgagg aaatagagcg ggtgatgcac
     1741 tgccttgaca aatccacaga agagcctatt gtgaaggtgg tggagaggga actcatttcc
     1801 aagcacatga agactattgt agaaatggaa aattctgggc tagtacatat gctgaaaaat
     1861 ggaaagacag aagaccttgc ttgcatgtac aaattattta gtcgtgtgcc aaatggtttg
     1921 aagaccatgt gtgaatgtat gagttgttat ttgagggaac aaggaaaagc ccttgtttct
     1981 gaggaaggag aagggaagaa tcctgttgac tatatccagg gcttattgga tctaaagagt
     2041 aggtttgatc gcttcctcca agaatccttc aataatgacc ggctctttaa acagactatt
     2101 gctggggact tcgagtattt cttaaacctc aactctaggt ctcctgaata cctctcatta
     2161 tttattgatg ataaactgaa aaagggagtc aaagggctaa cagaacaaga agtagaaaca
     2221 atattggata aggcaatggt cctttttagg tttatgcaag aaaaagatgt atttgaacgt
     2281 tattataaac aacacctggc aagaagactg ctcacaaata aaagtgtttc tgatgactct
     2341 gagaaaaata tgatttctaa attaaagact gagtgtggat gtcaattcac atcgaaactg
     2401 gaaggaatgt ttagggatat gagcatctca aacacaacta tggatgagtt caggcaacat
     2461 ctacaggcaa cgggggtctc tttaggtggt gttgatctca cagtccgggt tctcacaacg
     2521 ggatattggc ctactcagtc agccacacca aagtgcaaca tcccaccagc accaagacat
     2581 gcttttgaga tatttagaag gttctactta gccaaacaca gtggtcgaca gctcacactc
     2641 cagcatcaca tgggttctgc agatctcaat gccacctttt atggtccagt taaaaaggaa
     2701 gatggatctg aggttggtgt tggcggtgca caagtaactg gttctaatac acggaagcac
     2761 atactgcaag tctccacttt ccagatgacc atattaatgc tttttaataa tagagagaag
     2821 tacacatttg aggaaattca acaagagaca gatatccctg aaagagaact tgttagagcc
     2881 ctccagtccc tcgcctgtgg taaaccaaca cagcgggtcc tcacaaaaga acccaagtcc
     2941 aaggagatag aaagtggcca catatttaca gttaatgatc aattcacatc taaactacac
     3001 agagtcaaga ttcaaacagt tgcagccaaa caaggtgaat ccgacccaga aaggaaagaa
     3061 acaagacaga aagtagatga tgacagaaaa catgagatag aagctgctat agtgcgaata
     3121 atgaagtcta ggaagaagat gcagcacaat gttttagtag cagaggtaac tcagcaactg
     3181 aaggctcgat tcttaccaag tccagttgtt attaagaaac gtattgaagg acttattgag
     3241 agagaatatt tggcacgaac acctgaggat cgcaaagtat acacatatgt agcactcgag
     3301 cagaaactca tctcagaaga ggatctggca gcaaatgata tcctggatta caaggatgac
     3361 gacgataagg tttaaacggc cggccgcggt catagctgtt tcctgaacag atcccgggtg
     3421 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     3481 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     3541 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     3601 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     3661 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     3721 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     3781 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     3841 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     3901 aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca
     3961 accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca
     4021 gtagatgcct gttgaattgg gtacgcggcc agcggcgagc ggtatcagct cactcaaagg
     4081 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4141 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4201 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4261 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4321 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4381 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4441 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4501 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4561 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4621 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4681 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4741 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4801 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4861 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4921 tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg
     4981 ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc
     5041 cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt
     5101 tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     5161 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga
     5221 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     5281 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     5341 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
     5401 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     5461 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     5521 ctgatagcga tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     5581 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     5641 gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     5701 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     5761 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     5821 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     5881 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     5941 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
     6001 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     6061 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     6121 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     6181 aaacgatcct catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc
     6241 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     6301 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     6361 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     6421 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     6481 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     6541 cattccacag ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     6721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6901 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7021 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     7081 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt tt
//
