LOCUS       V037619                 7772 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037619
VERSION     V037619
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             908..3211
                     /label="CUL3(NM_003590)"
                     /note="CUL3(NM_003590)"
                     /gene="CUL3"
     CDS             3218..3244
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3248..3274
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3278..3304
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3339..3563
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3609..4037
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4051..4380
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4447..5241
                     /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    5415..5536
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5585..5601)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5609..5625
                     /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(5633..5663)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5678..5699
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5987..6572)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6743..7603)
                     /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(7604..7708)
                     /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 cgccaccatg tcgaatctga gcaaaggcac gggcagccgg aaggacacca agatgcggat
      961 ccgggccttt ccgatgacca tggatgaaaa atatgtaaac agcatttggg accttctgaa
     1021 aaatgcaatt caagaaatcc agcgtaagaa taacagtggt cttagttttg aggagctcta
     1081 tagaaatgca tatacaatgg ttttgcataa acatggagaa aagctctaca ctggactaag
     1141 agaagttgtt accgaacatc tcataaataa ggtgcgagaa gatgtactaa attcattgaa
     1201 taacaacttt cttcaaacgc taaatcaagc ttggaatgat catcaaacag ctatggtgat
     1261 gattagagac atactaatgt acatggaccg tgtgtatgta caacaaaata atgtggagaa
     1321 cgtctacaat ttgggattaa ttatttttcg agatcaagtt gtacgttatg ggtgtattag
     1381 ggatcatcta cggcaaactc tattggatat gattgcaaga gagcggaaag gagaagtcgt
     1441 agacagaggc gcaataagaa atgcttgcca gatgttaatg attttaggtc tcgaaggaag
     1501 atcagtctat gaagaagatt ttgaggctcc ttttttggaa atgtctgcag aattttttca
     1561 gatggaaagc cagaaatttt tagcagaaaa tagtgcttca gtatatataa agaaagtaga
     1621 agctagaatt aatgaagaaa tagaacgagt gatgcactgc cttgacaaat caacggaaga
     1681 accaattgta aaggtggttg aaagggaact catttccaag cacatgaaga ctatagtaga
     1741 aatggagaat tctgggctag tacatatgtt gaaaaatgga aagacagaag accttggttg
     1801 catgtacaag ttatttagtc gtgtgccaaa tggtttgaaa acaatgtgtg agtgtatgag
     1861 ttcctatttg agggagcaag gtaaagctct tgtttctgaa gaaggagaag gaaagaatcc
     1921 tgttgactat atccagggct tattggatct gaagagtagg ttcgatcgct tcctcctgga
     1981 atcattcaac aatgaccgtc tctttaaaca aactattgcg ggtgactttg agtattttct
     2041 caacctcaac tccaggtctc ctgaatacct ctcattattt attgatgata agctgaaaaa
     2101 gggagtcaaa gggctaacag aacaagaagt agaaacaata ttggataaag caatggtcct
     2161 ttttaggttt atgcaagaaa aagatgtatt tgaacgttat tataaacaac acttggcaag
     2221 gagacttctc acaaataaaa gtgtttctga tgactctgaa aaaaacatga tatctaagtt
     2281 aaagactgaa tgtggatgtc agttcacgtc aaaactggaa ggaatgttta gggatatgag
     2341 catctcaaac acaacgatgg atgaattcag gcaacatcta caggcaactg gtgtatcttt
     2401 aggtggtgtt gatcttacag tccgggtgct cacgacagga tattggccca ctcagtcagc
     2461 cacaccaaag tgcaacatcc caccagcacc aagacatgct tttgagatat tcagaaggtt
     2521 ctacttagcc aaacacagtg gtcgacagct cacactccag catcatatgg gttctgcaga
     2581 tctcaatgcc acattttatg gaccagttaa aaaggaagat ggatctgaag ttggtgttgg
     2641 aggtgcacaa gtaactggct ctaatacacg gaagcacata ttgcaagttt ccactttcca
     2701 gatgaccata ttaatgctct ttaataatag agaaaaatac acatttgagg aaattcagca
     2761 agagacagat atccctgaaa gagagcttgt tagagcccta cagtccctcg cctgtggtaa
     2821 accaacacag cgggttctta caaaagaacc caaatcaaag gaaatagaaa atggtcatat
     2881 atttacagtt aatgatcaat tcacatccaa actacacaga gtcaagattc aaacagttgc
     2941 tgccaaacaa ggtgaatccg acccagagag gaaagaaaca aggcagaaag tagacgacga
     3001 cagaaaacat gagatagaag ctgctatagt gcggataatg aaatctagaa agaagatgca
     3061 gcacaatgtt ctagtagcgg aggtaactca gcagttgaag gcgcgattct taccaagtcc
     3121 agttgttatt aagaaacgta ttgaaggact tattgagaga gaatatttgg cacgaacacc
     3181 tgaggatcgc aaagtataca catatgtagc actcgagtac ccatacgacg tacctgatta
     3241 tgcaacttat ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta
     3301 cgcatgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3361 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3421 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3481 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3541 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3601 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3661 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3721 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3781 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3841 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3901 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3961 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4021 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4141 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4201 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4261 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4321 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4381 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4441 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4501 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4561 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4621 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4681 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4741 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4801 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4861 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4921 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4981 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5041 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5101 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5161 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5221 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5281 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5341 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5401 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5521 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5581 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5641 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5761 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5821 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5881 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5941 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6001 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6061 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6121 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6181 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6241 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6301 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6361 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6421 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6481 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6541 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6601 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6661 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6721 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6781 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6841 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6901 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6961 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7021 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7081 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7141 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7201 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7261 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7321 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7381 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7441 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7501 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7561 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7621 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7681 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7741 acggatcggg agatctcccg atcccctatg gt
//
