LOCUS       V019196                 7139 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019196
VERSION     V019196
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             683..1819
                     /label="MAPK3(NM_002746)"
                     /note="MAPK3(NM_002746)"
                     /gene="MAPK3"
     CDS             1826..1891
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1915..2503
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2569..2617
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2819..3247
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3261..3590
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3657..4451
                     /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    4627..4748
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4797..4813)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4821..4837
                     /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(4845..4875)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4890..4911
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5199..5787)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5958..6818)
                     /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(6819..6923)
                     /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 ggatccgcca ccatggcggc ggcggcggct caggggggcg ggggcgggga
      721 gccccgtaga accgaggggg tcggcccggg ggtcccgggg gaggtggaga tggtgaaggg
      781 gcagccgttc gacgtgggcc cgcgctacac gcagttgcag tacatcggcg agggcgcgta
      841 cggcatggtc agctcggcct atgaccacgt gcgcaagact cgcgtggcca tcaagaagat
      901 cagccccttc gaacatcaga cctactgcca gcgcacgctc cgggagatcc agatcctgct
      961 gcgcttccgc catgagaatg tcatcggcat ccgagacatt ctgcgggcgt ccaccctgga
     1021 agccatgaga gatgtctaca ttgtgcagga cctgatggag actgacctgt acaagttgct
     1081 gaaaagccag cagctgagca atgaccatat ctgctacttc ctctaccaga tcctgcgggg
     1141 cctcaagtac atccactccg ccaacgtgct ccaccgagat ctaaagccct ccaacctgct
     1201 catcaacacc acctgcgacc ttaagatttg tgatttcggc ctggcccgga ttgccgatcc
     1261 tgagcatgac cacaccggct tcctgacgga gtatgtggct acgcgctggt accgggcccc
     1321 agagatcatg ctgaactcca agggctatac caagtccatc gacatctggt ctgtgggctg
     1381 cattctggct gagatgctct ctaaccggcc catcttccct ggcaagcact acctggatca
     1441 gctcaaccac attctgggca tcctgggctc cccatcccag gaggacctga attgtatcat
     1501 caacatgaag gcccgaaact acctacagtc tctgccctcc aagaccaagg tggcttgggc
     1561 caagcttttc cccaagtcag actccaaagc ccttgacctg ctggaccgga tgttaacctt
     1621 taaccccaat aaacggatca cagtggagga agcgctggct cacccctacc tggagcagta
     1681 ctatgacccg acggatgagc cagtggccga ggagcccttc accttcgcca tggagctgga
     1741 tgacctacct aaggagcggc tgaaggagct catcttccag gagacagcac gcttccagcc
     1801 cggagtgctg gaggcccccc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1861 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     1921 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1981 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2041 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2101 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2161 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2221 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2281 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2341 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2401 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2461 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2521 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2581 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2641 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2701 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2761 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2821 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2881 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2941 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     3001 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3061 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3121 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3181 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3241 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3301 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3361 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3421 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3481 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3541 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3601 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3661 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3721 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3781 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3841 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3901 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3961 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4021 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4081 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4141 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4201 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4261 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4321 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4381 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4441 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4501 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4561 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4621 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4681 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4741 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4801 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4861 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4921 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4981 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5041 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5101 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5161 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5221 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5281 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5341 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5401 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5461 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5521 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5581 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5641 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5701 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5761 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5821 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5881 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5941 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6001 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6061 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6121 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6181 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6241 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6301 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6361 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6421 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6481 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6541 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6601 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6661 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6721 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6781 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6841 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6901 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6961 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     7021 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     7081 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
