LOCUS       V043682                 7798 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043682
VERSION     V043682
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3234
                     /label="Cul3(NM_016716)"
                     /note="Cul3(NM_016716)"
                     /gene="Cul3"
     CDS             3241..3330
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3365..3589
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3635..4063
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4077..4406
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4473..5267
                     /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    5441..5562
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5611..5627)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5635..5651
                     /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(5659..5689)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5704..5725
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6013..6598)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6769..7629)
                     /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(7630..7734)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtcgaatc tgagcaaagg cacgggcagc
      961 cggaaggaca ccaagatgcg gatccgggcc tttccgatga ccatggatga aaaatatgta
     1021 aacagcattt gggaccttct gaaaaatgca attcaagaaa tccagcgtaa gaataacagt
     1081 ggtcttagtt ttgaggagct ctatagaaat gcatatacaa tggttttgca taaacatgga
     1141 gaaaagctct acactggact aagagaagtt gttactgaac atctcataaa taaggtgcga
     1201 gaagatgtac taaattcatt gaataacaac tttcttcaaa cactaaatca agcttggaat
     1261 gatcatcaaa cagccatggt gatgattaga gacatattaa tgtatatgga ccgtgtatat
     1321 gtacaacaga ataatgtaga aaatgtctac aatttgggat taataatttt tcgagatcaa
     1381 gttgtacggt atgggtgtat tagggatcat cttcggcaaa ctttattgga tatgattgcc
     1441 agagagcgga aaggagaagt tgtagacaga ggagcaataa gaaatgcttg ccagatgtta
     1501 atgattttag gtcttgaagg aagatcagtc tatgaggaag attttgaggc tccgtttttg
     1561 gaaatgtctg cagaattttt tcagatggaa agccagaaat ttttagcaga aaacagtgct
     1621 tcagtatata taaagaaagt agaagctagg attaatgagg aaatagagcg ggtgatgcac
     1681 tgccttgaca aatccacaga agagcctatt gtgaaggtgg tggagaggga actcatttcc
     1741 aagcacatga agactattgt agaaatggaa aattctgggc tagtacatat gctgaaaaat
     1801 ggaaagacag aagaccttgc ttgcatgtac aaattattta gtcgtgtgcc aaatggtttg
     1861 aagaccatgt gtgaatgtat gagttgttat ttgagggaac aaggaaaagc ccttgtttct
     1921 gaggaaggag aagggaagaa tcctgttgac tatatccagg gcttattgga tctaaagagt
     1981 aggtttgatc gcttcctcca agaatccttc aataatgacc ggctctttaa acagactatt
     2041 gctggggact tcgagtattt cttaaacctc aactctaggt ctcctgaata cctctcatta
     2101 tttattgatg ataaactgaa aaagggagtc aaagggctaa cagaacaaga agtagaaaca
     2161 atattggata aggcaatggt cctttttagg tttatgcaag aaaaagatgt atttgaacgt
     2221 tattataaac aacacctggc aagaagactg ctcacaaata aaagtgtttc tgatgactct
     2281 gagaaaaata tgatttctaa attaaagact gagtgtggat gtcaattcac atcgaaactg
     2341 gaaggaatgt ttagggatat gagcatctca aacacaacta tggatgagtt caggcaacat
     2401 ctacaggcaa cgggggtctc tttaggtggt gttgatctca cagtccgggt tctcacaacg
     2461 ggatattggc ctactcagtc agccacacca aagtgcaaca tcccaccagc accaagacat
     2521 gcttttgaga tatttagaag gttctactta gccaaacaca gtggtcgaca gctcacactc
     2581 cagcatcaca tgggttctgc agatctcaat gccacctttt atggtccagt taaaaaggaa
     2641 gatggatctg aggttggtgt tggcggtgca caagtaactg gttctaatac acggaagcac
     2701 atactgcaag tctccacttt ccagatgacc atattaatgc tttttaataa tagagagaag
     2761 tacacatttg aggaaattca acaagagaca gatatccctg aaagagaact tgttagagcc
     2821 ctccagtccc tcgcctgtgg taaaccaaca cagcgggtcc tcacaaaaga acccaagtcc
     2881 aaggagatag aaagtggcca catatttaca gttaatgatc aattcacatc taaactacac
     2941 agagtcaaga ttcaaacagt tgcagccaaa caaggtgaat ccgacccaga aaggaaagaa
     3001 acaagacaga aagtagatga tgacagaaaa catgagatag aagctgctat agtgcgaata
     3061 atgaagtcta ggaagaagat gcagcacaat gttttagtag cagaggtaac tcagcaactg
     3121 aaggctcgat tcttaccaag tccagttgtt attaagaaac gtattgaagg acttattgag
     3181 agagaatatt tggcacgaac acctgaggat cgcaaagtat acacatatgt agcactcgag
     3241 gaacaaaaac taatatcaga ggaagatttg gaacagaaac taatcagtga agaagattta
     3301 gagcaaaagt taatttccga ggaggactta tgagggcccg tttaaacccg ctgatcagcc
     3361 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3421 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3481 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3541 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3601 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3841 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3961 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4081 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4141 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4201 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4261 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4321 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4381 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4441 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4501 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4561 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4621 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4681 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4741 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4801 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4861 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4921 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4981 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5041 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5101 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5161 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5221 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5281 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5341 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5401 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5461 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5521 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5581 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5641 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5701 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5761 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5821 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5881 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5941 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6001 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6061 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6181 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6421 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6541 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6901 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7741 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggt
//
