LOCUS       V043830                 7591 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043830
VERSION     V043830
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1033..3111
                     /label="Tab2(NM_138667)"
                     /note="Tab2(NM_138667)"
                     /gene="Tab2"
     polyA_signal    3191..3415
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3461..3889
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3903..4232
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4299..5093
                     /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    5267..5388
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5437..5453)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5461..5477
                     /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(5485..5515)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5530..5551
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5839..6424)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6595..7455)
                     /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(7456..7560)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttatggccca aggaagccac caaattgatt ttcaggtttt acatgacctg
     1081 cgacaaaaat tcccagaagt gcctgaagtt gttgtatcca ggtgcatgtt acagaacaat
     1141 aataacctgg atgcctgctg cgctgttctc tctcaggaga gtacaaggta tctttatggt
     1201 gaaggagacc tgaatttttc agatgagtct ggaatttctg gtctacgcaa tcacatgact
     1261 tctctcaact tggacttgca gtcacagaat gtttaccacc atggaagaga aggaagtcga
     1321 gtgaatggaa gcaggacttt aacacacagc gttagtgatg gacagcttca cggtgggcag
     1381 tccaataacg aactgtttca gcaagagcca cagacagcac cagctcaagt tcctcaaggc
     1441 tttaatgttt ttggaatgcc cagtacatct ggtgcttcaa actcaacacc gcatcttgga
     1501 tttcacctag gcagcaaagg gacatcaaac ctttctcaac agactcccag gttcaatcct
     1561 attatggtaa ctttagcccc aaacatacag actggccgta gtactcctac atctttgcac
     1621 atacatggtg tacctccgcc tgtactgaac agtccacagg gaaattctat ctatattagg
     1681 ccttatatta caactcctag tggtacagct cggcagacac aacaacattc tggctgggtg
     1741 tctcagttta atcccatgaa ccctcagcaa gcctatcaac cttcacagcc tggaccttgg
     1801 actacttatc ctgcatccaa tcctttgcca catacctcaa cccagcagcc gaatcagcag
     1861 ggccatcaga cctctcatgt ctacatgcct atcagttcgc caactactcc acaaccgcca
     1921 accattcact catctgggag ctctcagtct tctgcccata gccaatataa cattcagaat
     1981 atttcaacag gacctcgaaa gaaccagata gaaatcaaac ttgaaccccc acaaagaaac
     2041 agttcttcaa aattacgttc ttctggacct cgaactgcca gcacttcctc attggtcaac
     2101 agccagacct taaatagaaa tcagcccact gtttacatag ctgccagtcc tccaaatact
     2161 gatgagatga tctcccgtag tcaacccaag gtctatattt cagccaatgc caccgcagga
     2221 gatgagcaag gcatgcggaa tcaaccgaca ctcttcatat ctacaaactc tgggccatct
     2281 gcagcctcca ggaacatgtc tgggcaagtg agcatgggtc ctgcctttat ccatcaccat
     2341 ccacccaaaa gccgagtgtt aggtggtaac tctgcaacgt ctcctcgagt ggtggtcact
     2401 caacccaaca caaaatatac tttcaaaatt acagtttccc ccaataagcc ccctgctgtt
     2461 tccccggggg tggtctcccc aacctttgaa cttacaaatc ttctaaatca tcctgaccat
     2521 tatgtagaaa cagagaacat tcagcatctc acagacccgg ctctagcaca tgtggataga
     2581 ataagcgaag cccggaaact gagtatggga tctgatgatg ctgcctacac acaagctctg
     2641 ctggtgcacc agaaggccag gatggaacgg cttcaaagag agctcgagat gcaaaagaaa
     2701 aagctggata aactcaaatc tgaggtcaat gagatggaaa ataatctaac tcgaaggcgc
     2761 ctgaagagat caaattccat ttcccagata ccgtcactcg aagaaatgca gcagttgaga
     2821 agttgtaata gacaactcca gattgacatt gactgcttaa ccaaagaaat tgatcttttt
     2881 caagcccgag gaccacattt taaccccagc gctattcata acttttatga caatattgga
     2941 tttgtaggcc ctgtgccacc aaaacccaaa gatcaaaggt ccaccatcaa agcaccaaag
     3001 acccaagacg cagaggatga ggaaggtgct cagtggaatt gcactgcctg tacgtttctg
     3061 aatcacccgg ccttaatccg ctgtgaacag tgtgagatgc ctcggcattt ctgagaattc
     3121 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     3181 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3241 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3301 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3361 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3421 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3481 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3541 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3601 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3661 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3721 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3781 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3841 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3901 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3961 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4021 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4081 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4141 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4201 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4261 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4321 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4381 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4441 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4501 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4561 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4621 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4681 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4741 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4801 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4861 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4921 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4981 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5041 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5101 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5161 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5221 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5281 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5341 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5401 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5461 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5521 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5581 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6241 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6361 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6421 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6481 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6541 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6601 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6661 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6721 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6781 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6841 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6901 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6961 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7021 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7081 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7141 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7201 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7261 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7321 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7381 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7441 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7501 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7561 cacatttccc cgaaaagtgc cacctgacgt c
//
