LOCUS       V022494                 7000 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022494
VERSION     V022494
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"
     CDS             925..2739
                     /label="MAP3K7(NM_145331)"
                     /note="MAP3K7(NM_145331)"
                     /gene="MAP3K7"
     CDS             2764..2829
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2864..3088
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3134..3562
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3576..3905
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3917..4516
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    4676..4797
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4846..4862)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4870..4886
                     /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(4894..4924)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4939..4960
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5248..5833)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6004..6864)
                     /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(6865..6969)
                     /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 aagctttcta gagaattcgc caccatgtct acagcctctg ccgcctcctc ctcctcctcg
      961 tcttcggccg gtgagatgat cgaagcccct tcccaggtcc tcaactttga agagatcgac
     1021 tacaaggaga tcgaggtgga agaggttgtt ggaagaggag cctttggagt ttgcaaagct
     1081 aagtggagag caaaagatgt tgctattaaa caaatagaaa gtgaatctga gaggaaagcg
     1141 tttattgtag agcttcggca gttatcccgt gtgaaccatc ctaatattgt aaagctttat
     1201 ggagcctgct tgaatccagt gtgtcttgtg atggaatatg ctgaaggggg ctctttatat
     1261 aatgtgctgc atggtgctga accattgcca tattatactg ctgcccacgc aatgagttgg
     1321 tgtttacagt gttcccaagg agtggcttat cttcacagca tgcaacccaa agcgctaatt
     1381 cacagggacc tgaaaccacc aaacttactg ctggttgcag gggggacagt tctaaaaatt
     1441 tgtgattttg gtacagcctg tgacattcag acacacatga ccaataacaa ggggagtgct
     1501 gcttggatgg cacctgaagt ttttgaaggt agtaattaca gtgaaaaatg tgacgtcttc
     1561 agctggggta ttattctttg ggaagtgata acgcgtcgga aaccctttga tgagattggt
     1621 ggcccagctt tccgaatcat gtgggctgtt cataatggta ctcgaccacc actgataaaa
     1681 aatttaccta agcccattga gagcctgatg actcgttgtt ggtctaaaga tccttcccag
     1741 cgcccttcaa tggaggaaat tgtgaaaata atgactcact tgatgcggta ctttccagga
     1801 gcagatgagc cattacagta tccttgtcag tattcagatg aaggacagag caactctgcc
     1861 accagtacag gctcattcat ggacattgct tctacaaata cgagtaacaa aagtgacact
     1921 aatatggagc aagttcctgc cacaaatgat actattaagc gcttagaatc aaaattgttg
     1981 aaaaatcagg caaagcaaca gagtgaatct ggacgtttaa gcttgggagc ctcccgtggg
     2041 agcagtgtgg agagcttgcc cccaacctct gagggcaaga ggatgagtgc tgacatgtct
     2101 gaaatagaag ctaggatcgc cgcaaccaca gcctattcca agcctaaacg gggccaccgt
     2161 aaaactgctt catttggcaa cattctggat gtccctgaga tcgtcatatc aggcaacgga
     2221 cagccaagac gtagatccat ccaagacttg actgtaactg gaacagaacc tggtcaggtg
     2281 agcagtaggt catccagtcc cagtgtcaga atgattacta cctcaggacc aacctcagaa
     2341 aagccaactc gaagtcatcc atggacccct gatgattcca cagataccaa tggatcagat
     2401 aactccatcc caatggctta tcttacactg gatcaccaac tacagcctct agcaccgtgc
     2461 ccaaactcca aagaatctat ggcagtgttt gaacagcatt gtaaaatggc acaagaatat
     2521 atgaaagttc aaacagaaat tgcattgtta ttacagagaa agcaagaact agttgcagaa
     2581 ctggaccagg atgaaaagga ccagcaaaat acatctcgcc tggtacagga acataaaaag
     2641 cttttagatg aaaacaaaag cctttctact tactaccagc aatgcaaaaa acaactagag
     2701 gtcatcagaa gtcagcagca gaaacgacaa ggcacttcag gatccggtac cctcgagacc
     2761 ggtgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
     2821 gatgacaagt gaccggtaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc
     2881 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2941 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3001 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3061 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3121 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3181 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3241 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3301 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3361 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3421 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3481 tcttttgatt tataagggat tttggccatt tcggcctatt ggttaaaaaa tgagctgatt
     3541 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3601 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3661 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3721 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3781 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3841 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3901 gcaaaaagct cccgggatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3961 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4021 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4081 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4141 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4201 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     4261 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     4321 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4381 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4441 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4501 caagcccggt gcctgattcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4561 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4621 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4681 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4741 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4801 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4861 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4921 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4981 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5041 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5101 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5761 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6001 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6061 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6121 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6181 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6241 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6301 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6361 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6421 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6481 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6541 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6601 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6661 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6721 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6781 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6841 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6901 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6961 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
