LOCUS       V023857                 7379 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023857
VERSION     V023857
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             779..2059
                     /label="MAPK8(NM_001323330)"
                     /note="MAPK8(NM_001323330)"
                     /gene="MAPK8"
     CDS             2066..2131
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2155..2743
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2809..2857
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3059..3487
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3501..3830
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3897..4691
                     /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    4867..4988
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5037..5053)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5061..5077
                     /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(5085..5115)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5130..5151
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5439..6027)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6198..7058)
                     /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(7059..7163)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 gagcagaagc aagcgtgaca acaattttta tagtgtagag attggagatt ctacattcac
      841 agtcctgaaa cgatatcaga atttaaaacc tataggctca ggagctcaag gaatagtatg
      901 cgcagcttat gatgccattc ttgaaagaaa tgttgcaatc aagaagctaa gccgaccatt
      961 tcagaatcag actcatgcca agcgggccta cagagagcta gttcttatga aatgtgttaa
     1021 tcacaaaaat ataattggcc ttttgaatgt tttcacacca cagaaatccc tagaagaatt
     1081 tcaagatgtt tacatagtca tggagctcat ggatgcaaat ctttgccaag tgattcagat
     1141 ggagctagat catgaaagaa tgtcctacct tctctatcag atgctgtgtg gaatcaagca
     1201 ccttcattct gctggaatta ttcatcggga cttaaagccc agtaatatag tagtaaaatc
     1261 tgattgcact ttgaagattc ttgacttcgg tctggccagg actgcaggaa cgagttttat
     1321 gatgacgcct tatgtagtga ctcgctacta cagagcaccc gaggtcatcc ttggcatggg
     1381 ctacaaggaa aacgttgaca tttggtcagt tgggtgcatc atgggagaaa tgatcaaagg
     1441 tggtgttttg ttcccaggta cagatcatat tgatcagtgg aataaagtta ttgaacagct
     1501 tggaacacca tgtcctgaat tcatgaagaa actgcaacca acagtaagga cttacgttga
     1561 aaacagacct aaatatgctg gatatagctt tgagaaactc ttccctgatg tccttttccc
     1621 agctgactca gaacacaaca aacttaaagc cagtcaggca agggatttgt tatccaaaat
     1681 gctggtaata gatgcatcta aaaggatctc tgtagatgaa gctctccaac acccgtacat
     1741 caatgtctgg tatgatcctt ctgaagcaga agctccacca ccaaagatcc ctgacaagca
     1801 gttagatgaa agggaacaca caatagaaga gtggaaagaa ttgatatata aggaagttat
     1861 ggacttggag gagagaacca agaatggagt tatacggggg cagccctctc ctttaggtgc
     1921 agcagtgatc aatggctctc agcatccatc atcatcgtcg tctgtcaatg atgtgtcttc
     1981 aatgtcaaca gatccgactt tggcctctga tacagacagc agtctagaag cagcagctgg
     2041 gcctctgggc tgctgtagac tcgaggacta caaagaccat gacggtgatt ataaagatca
     2101 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     2161 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2221 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2281 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2341 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2401 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2461 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2521 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2581 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2641 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2701 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2761 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2821 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2881 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2941 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     3001 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     3061 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3121 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3181 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     3241 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3301 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3361 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3421 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3481 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3661 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3721 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3781 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3841 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3901 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3961 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4021 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4081 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4141 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4201 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4261 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4321 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4381 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4441 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4501 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4561 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4621 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4681 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4741 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4801 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4861 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4921 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4981 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5041 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5101 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5161 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5221 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5281 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5341 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5401 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5461 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5521 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5581 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5641 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5701 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5761 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5821 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5881 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5941 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6001 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6481 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6541 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6601 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6661 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6721 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6781 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6841 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6901 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6961 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7021 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7081 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7141 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7201 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     7261 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     7321 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
