LOCUS       V032541                 6722 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032541
VERSION     V032541
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..2188
                     /label="MAPK8(NM_139049)"
                     /note="MAPK8(NM_139049)"
                     /gene="MAPK8"
     CDS             2189..2215
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2222..2251
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     polyA_signal    2289..2513
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2559..2987
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3001..3330
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3397..4191
                     /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    4365..4486
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4535..4551)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4559..4575
                     /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(4583..4613)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4628..4649
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4937..5522)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5693..6553)
                     /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(6554..6658)
                     /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 agcagaagca agcgtgacaa caatttttat agtgtagaga ttggagattc
      961 tacattcaca gtcctgaaac gatatcagaa tttaaaacct ataggctcag gagctcaagg
     1021 aatagtatgc gcagcttatg atgccattct tgaaagaaat gttgcaatca agaagctaag
     1081 ccgaccattt cagaatcaga ctcatgccaa gcgggcctac agagagctag ttcttatgaa
     1141 atgtgttaat cacaaaaata taattggcct tttgaatgtt ttcacaccac agaaatccct
     1201 agaagaattt caagatgttt acatagtcat ggagctcatg gatgcaaatc tttgccaagt
     1261 gattcagatg gagctagatc atgaaagaat gtcctacctt ctctatcaga tgctgtgtgg
     1321 aatcaagcac cttcattctg ctggaattat tcatcgggac ttaaagccca gtaatatagt
     1381 agtaaaatct gattgcactt tgaagattct tgacttcggt ctggccagga ctgcaggaac
     1441 gagttttatg atgacgcctt atgtagtgac tcgctactac agagcacccg aggtcatcct
     1501 tggcatgggc tacaaggaaa acgtggattt atggtctgtg gggtgcatta tgggagaaat
     1561 ggtttgccac aaaatcctct ttccaggaag ggactatatt gatcagtgga ataaagttat
     1621 tgaacagctt ggaacaccat gtcctgaatt catgaagaaa ctgcaaccaa cagtaaggac
     1681 ttacgttgaa aacagaccta aatatgctgg atatagcttt gagaaactct tccctgatgt
     1741 ccttttccca gctgactcag aacacaacaa acttaaagcc agtcaggcaa gggatttgtt
     1801 atccaaaatg ctggtaatag atgcatctaa aaggatctct gtagatgaag ctctccaaca
     1861 cccgtacatc aatgtctggt atgatccttc tgaagcagaa gctccaccac caaagatccc
     1921 tgacaagcag ttagatgaaa gggaacacac aatagaagag tggaaagaat tgatatataa
     1981 ggaagttatg gacttggagg agagaaccaa gaatggagtt atacgggggc agccctctcc
     2041 tttaggtgca gcagtgatca atggctctca gcatccatca tcatcgtcgt ctgtcaatga
     2101 tgtgtcttca atgtcaacag atccgacttt ggcctctgat acagacagca gtctagaagc
     2161 agcagctggg cctctgggct gctgtagata cccatacgat gttccagatt acgctctcga
     2221 ggagcagaag ctgatctcag aggaggacct gtagtgaggg cccgtttaaa cccgctgatc
     2281 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2341 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2401 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2461 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2521 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2581 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2641 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2701 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2761 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2821 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2881 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2941 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3001 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3061 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3301 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3361 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3421 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3481 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3541 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3601 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3661 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3721 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3781 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3841 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3901 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3961 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4021 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4081 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4141 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4201 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4261 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4321 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4381 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4441 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4501 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4561 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4621 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4681 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4741 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4801 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4861 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4921 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4981 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5041 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5101 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5161 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5221 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5281 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5341 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5401 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5461 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6301 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6661 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6721 gt
//
