LOCUS       V040439                 7450 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040439
VERSION     V040439
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"
     CDS             706..2106
                     /label="MAP3K8(NM_005204)"
                     /note="MAP3K8(NM_005204)"
                     /gene="MAP3K8"
     CDS             2113..2202
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2226..2814
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2880..2928
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3130..3558
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3572..3901
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3968..4762
                     /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    4938..5059
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5108..5124)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5132..5148
                     /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(5156..5186)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5201..5222
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5510..6098)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6269..7129)
                     /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(7130..7234)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga gtacatgagc
      721 actggaagtg acaataaaga agagattgat ttattaatta aacatttaaa tgtgtctgat
      781 gtaatagaca ttatggaaaa tctttatgca agtgaagagc cagcagttta tgaacccagt
      841 ctaatgacca tgtgtcaaga cagtaatcaa aacgatgagc gttctaagtc tctgctgctt
      901 agtggccaag aggtaccatg gttgtcatca gtcagatatg gaactgtgga ggatttgctt
      961 gcttttgcaa accatatatc caacactgca aagcattttt atggacaacg accacaggaa
     1021 tctggaattt tattaaacat ggtcatcact ccccaaaatg gacgttacca aatagattcc
     1081 gatgttctcc tgatcccctg gaagctgact tacaggaata ttggttctga ttttattcct
     1141 cggggcgcct ttggaaaggt atacttggca caagatataa agacgaagaa aagaatggcg
     1201 tgtaaactga tcccagtaga tcaatttaag ccatctgatg tggaaatcca ggcttgcttc
     1261 cggcacgaga acatcgcaga gctgtatggc gcagtcctgt ggggtgaaac tgtccatctc
     1321 tttatggaag caggcgaggg agggtctgtt ctggagaaac tggagagctg tggaccaatg
     1381 agagaatttg aaattatttg ggtgacaaag catgttctca agggacttga ttttctacac
     1441 tcaaagaaag tgatccatca tgatattaaa cctagcaaca ttgttttcat gtccacaaaa
     1501 gctgttttgg tggattttgg cctaagtgtt caaatgaccg aagatgtcta ttttcctaag
     1561 gacctccgag gaacagagat ttacatgagc ccagaggtca tcctgtgcag gggccattca
     1621 accaaagcag acatctacag cctgggggcc acgctcatcc acatgcagac gggcacccca
     1681 ccctgggtga agcgctaccc tcgctcagcc tatccctcct acctgtacat aatccacaag
     1741 caagcacctc cactggaaga cattgcagat gactgcagtc cagggatgag agagctgata
     1801 gaagcttccc tggagagaaa ccccaatcac cgcccaagag ccgcagacct actaaaacat
     1861 gaggccctga acccgcccag agaggatcag ccacgctgtc agagtctgga ctctgccctc
     1921 ttggagcgca agaggctgct gagtaggaag gagctggaac ttcctgagaa cattgctgat
     1981 tcttcgtgca caggaagcac cgaggaatct gagatgctca agaggcaacg ctctctctac
     2041 atcgacctcg gcgctctggc tggctacttc aatcttgttc ggggaccacc aacgcttgaa
     2101 tatggcctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     2161 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc
     2221 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2281 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2341 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2401 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2461 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2521 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2581 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     2641 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2701 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2761 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     2821 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     2881 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     2941 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3001 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3061 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     3121 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3181 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3241 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3301 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3361 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3421 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3481 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     3541 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3721 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3841 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3901 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3961 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4021 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4081 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4141 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4201 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4261 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4321 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4381 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4441 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4501 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4561 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4621 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4681 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4741 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     4801 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4921 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4981 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5041 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5101 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5701 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5941 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7261 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     7321 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     7381 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     7441 tctgcttagg
//
