LOCUS       V045748                 8235 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045748
VERSION     V045748
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2915
                     /label="Mapk6(NM_015806)"
                     /note="Mapk6(NM_015806)"
                     /gene="Mapk6"
     CDS             2922..2987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3011..3599
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3665..3713
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3915..4343
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4357..4686
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4753..5547
                     /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    5723..5844
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5893..5909)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5917..5933
                     /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(5941..5971)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5986..6007
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6295..6883)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7054..7914)
                     /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(7915..8019)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc agagaaattc gaaagtctca
      781 tgaacattca tggctttgat ctgggctcta ggtacatgga cttaaaacca ttgggctgcg
      841 gaggcaatgg cttggtcttt tctgctgtag acaatgactg tgacaaaaga gtagccatca
      901 agaaaattgt cctcaccgat ccccagagtg tcaaacatgc tctccgtgaa atcaaaatta
      961 ttcgaagact tgaccacgat aacattgtga aagtgtttga aattcttggt cccagtggaa
     1021 gccagttaac agacgatgtg ggctctctca cggagctgaa tagcgtctac attgttcagg
     1081 agtatatgga gacagacttg gcgaatgtgc tggagcaggg ccctttactg gaagagcatg
     1141 ccaggctttt catgtatcag ctgctacgtg ggctcaaata tatccattct gcaaacgtac
     1201 tgcacagaga tctcaagcca gctaatcttt tcattaacac tgaagacttg gtgctgaaga
     1261 taggtgactt tggcctggca cggatcatgg atcctcatta ttcccataag ggtcatcttt
     1321 ctgaaggatt ggttaccaaa tggtacagat ctccacggct tttactttct cctaataatt
     1381 acactaaagc cattgacatg tgggctgcag gctgcatctt tgctgagatg ctgactggta
     1441 aaaccctctt tgcaggtgca catgaacttg aacagatgca gctgatcctc gactccatcc
     1501 ctgttgtgca cgaggaggat cggcaggagc ttctcagcgt gattccagtt tacattagaa
     1561 acgacatgac tgagccacac agaccgctca ctcagctgct gccggggatc agccgagaag
     1621 cactggattt cctggagcag attctgactt tcagccccat ggaccggctg acagctgagg
     1681 aagccctttc ccatccttac atgagcatct actccttccc gacggatgag cccatctcaa
     1741 gccacccttt ccacatagag gacgaagtgg acgacatttt gcttatggat gaaacacaca
     1801 gtcacattta taactgggaa aggtaccacg attgtcagtt ctcggagcat gactggccta
     1861 ttcataacaa ctttgatatc gatgaggtgc agcttgaccc aagagctctg tctgacgtca
     1921 ccgatgaaga agaagtccaa gttgatcctc gaaaatactt ggatggagac cgagagaagt
     1981 atctagagga tcccgccttc gacaccagct actctgctga accctgctgg cagtacccag
     2041 atcaccacga gaacaagtac tgcgatctgg agtgtagcca cacctgtaac tacaaaacaa
     2101 gatcatcacc atacttagat aacctggtgt ggagggagag cgaggttaac cattactatg
     2161 agcccaagct tattatagat ctttccaact ggaaagagca aagtaaagaa aaatccgata
     2221 agagaggcaa gtccaagtgt gagaggaacg ggttggtcaa ggcgcagatc gcgctagagg
     2281 aagcatccca gcagctggct gagagggaga ggggccaagg cttcgacttt gactccttca
     2341 tcgcggggac cattcagctc agtgcccagc atcagtctgc tgacgtagtt gacaagttaa
     2401 acgacttgaa tagctcagtg tcccagctag aattgaaaag cctgatatcc aagtcagtca
     2461 gccgagaaaa gcaagaaaag ggaagggcta acctggccca gctgggagcc ttgtaccagt
     2521 cctcctggga cagccagttt gtgagtggcg gggaggagtg cttcctcatc agtcagtttt
     2581 gttgtgaggt caggaaggac gagcatgcgg agaaggagaa cacctacacc agctatttgg
     2641 acaagttttt tagcaggaag gaggattccg aaatgctaga aactgagcca gtggaggaag
     2701 ggaagcgtgg ggagagaggc cgtgaggcag ggcttctgag cggcggtggg gagttcctcc
     2761 tgagcaagca gctggagtcc ataggcaccc cgcagttcca cagcccagtg gggtccccac
     2821 tcaagtccat ccaggccacc ttgacacctt ccgctatgaa atcttcccct caaatccctc
     2881 acaagacata cagcagcatt ctgaaacatc tgaacggatc cgactacaaa gaccatgacg
     2941 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     3001 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3061 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3121 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3181 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3241 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3301 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3361 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3421 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3481 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3541 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3601 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3661 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     3721 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     3781 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3841 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     3901 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3961 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4021 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     4081 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4141 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4201 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4261 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     4321 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4381 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4441 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4501 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4561 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4621 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4681 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4741 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4801 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4861 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4921 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4981 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5041 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5101 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5161 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5221 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5281 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5341 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5401 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5461 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5521 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     5581 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5641 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5701 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5761 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5821 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5881 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5941 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6001 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6061 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6121 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6181 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6241 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6301 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6361 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6421 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6481 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6541 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6601 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6661 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6721 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6781 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6841 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6901 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6961 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7021 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7081 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7141 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7201 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7261 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7321 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7381 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7441 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7501 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7561 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7621 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7681 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7741 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7801 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7861 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7921 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7981 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8041 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     8101 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     8161 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     8221 aagaatctgc ttagg
//
