LOCUS       V027535                 8057 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027535
VERSION     V027535
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             941..3547
                     /label="MUSK(NM_005592)"
                     /note="MUSK(NM_005592)"
                     /gene="MUSK"
     CDS             3554..3619
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3657..3881
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3927..4355
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4369..4698
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4765..5559
                     /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    5733..5854
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5903..5919)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5927..5943
                     /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(5951..5981)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5996..6017
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6305..6890)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7061..7921)
                     /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(7922..8026)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgagagagc tcgtcaacat
      961 tccactggta catattctta ctctggttgc cttcagcgga actgagaaac ttccaaaagc
     1021 tcctgtcatc accactcctc ttgaaacagt ggatgcctta gttgaagaag tggctacttt
     1081 catgtgtgca gtggaatcct acccccagcc tgagatttcc tggactagaa ataaaattct
     1141 cattaaactc tttgacaccc ggtacagcat ccgggagaat gggcagctcc tcaccatcct
     1201 gagtgtggaa gacagtgatg atggcattta ctgctgcacg gccaacaatg gtgtgggagg
     1261 agctgtggag agttgtggag ccctgcaagt gaagatgaaa cctaaaataa ctcgtcctcc
     1321 cataaatgtg aaaataatag agggattaaa agcagtccta ccatgtacta caatgggtaa
     1381 tcccaaacca tcagtgtctt ggataaaggg agacagccct ctcagggaaa attcccgaat
     1441 tgcagttctt gaatctggga gcttgaggat tcataacgta caaaaggaag atgcaggaca
     1501 gtatcgatgt gtggcaaaaa acagcctcgg gacagcatat tccaaagtgg tgaagctgga
     1561 agttgaggtt tttgccagga tcctgcgggc tcctgaatcc cacaatgtca cctttggctc
     1621 ctttgtgacc ctgcactgta cagcaacagg cattcctgtc cccaccatca cctggattga
     1681 aaacggaaat gctgtttctt ctgggtccat tcaagagagt gtgaaagacc gagtgattga
     1741 ctcaagactg cagctgttta tcaccaagcc aggactctac acatgcatag ctaccaataa
     1801 gcatggggag aagttcagta ctgccaaggc tgcagccacc atcagcatag cagaatggag
     1861 taaaccacag aaagataaca aaggctactg cgcccagtac agaggggagg tgtgtaatgc
     1921 agtcctggca aaagatgctc ttgtttttct caacacctcc tatgcggacc ctgaggaggc
     1981 ccaagagcta ctggtccaca cggcctggaa tgaactgaaa gtagtgagcc cagtctgccg
     2041 gccagctgct gaggctttgt tgtgtaacca catcttccag gagtgcagtc ctggagtagt
     2101 gcctactcct attcccattt gcagagagta ctgcttggca gtaaaggagc tcttctgcgc
     2161 aaaagaatgg ctggtaatgg aagagaagac ccacagagga ctctacagat ccgagatgca
     2221 tttgctgtcc gtgccagaat gcagcaagct tcccagcatg cattgggacc ccacggcctg
     2281 tgccagactg ccacatctag attataacaa agaaaaccta aaaacattcc caccaatgac
     2341 gtcctcaaag ccaagtgtgg acattccaaa tctgccttcc tcctcctctt cttccttctc
     2401 tgtctcacct acatactcca tgactgtaat aatctccatc atgtccagct ttgcaatatt
     2461 tgtgcttctt accataacta ctctctattg ctgccgaaga agaaaacaat ggaaaaataa
     2521 gaaaagagaa tcagcagcag taaccctcac cacactgcct tctgagctct tactagatag
     2581 acttcatccc aaccccatgt accagaggat gccgctcctt ctgaacccca aattgctcag
     2641 cctggagtat ccaaggaata acattgaata tgtgagagac atcggagagg gagcgtttgg
     2701 aagggtgttt caagcaaggg caccaggctt acttccctat gaacctttca ctatggtggc
     2761 agtaaagatg ctcaaagaag aagcctcggc agatatgcaa gcggactttc agagggaggc
     2821 agccctcatg gcagaatttg acaaccctaa cattgtgaag ctattaggag tgtgtgctgt
     2881 cgggaagcca atgtgcctgc tctttgaata catggcctat ggtgacctca atgagttcct
     2941 ccgcagcatg tcccctcaca ccgtgtgcag cctcagtcac agtgacttgt ctatgagggc
     3001 tcaggtctcc agccctgggc ccccacccct ctcctgtgct gagcagcttt gcattgccag
     3061 gcaggtggca gctggcatgg cttacctctc agaacgtaag tttgttcacc gagatttagc
     3121 caccaggaac tgcctggtgg gcgagaacat ggtggtgaaa attgccgact ttggcctctc
     3181 caggaacatc tactcagcag actactacaa agctaatgaa aacgacgcta tccctatccg
     3241 ttggatgcca ccagagtcca ttttttataa ccgctacact acagagtctg atgtgtgggc
     3301 ctatggcgtg gtcctctggg agatcttctc ctatggcctg cagccctact atgggatggc
     3361 ccatgaggag gtcatttact acgtgcgaga tggcaacatc ctctcctgcc ctgagaactg
     3421 ccccgtggag ctgtacaatc tcatgcgtct atgttggagc aagctgcctg cagacagacc
     3481 cagtttcacc agtattcacc gaattctgga acgcatgtgt gagagggcag agggaactgt
     3541 gagtgtcctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     3601 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt
     3661 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3721 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3781 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3841 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3901 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3961 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4021 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4081 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4141 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4201 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4261 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4321 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4381 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4441 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4501 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4561 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4621 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4681 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4741 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4801 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4861 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4921 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4981 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5041 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5101 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5161 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5221 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5281 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5341 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5401 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5461 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5521 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5581 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5641 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5701 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5761 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5821 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5881 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5941 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6001 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6061 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6121 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6181 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6241 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6301 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6361 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6421 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6481 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6541 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6601 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6661 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6721 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6781 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6841 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6901 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6961 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7021 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7081 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7141 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7201 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7261 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7321 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7381 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7441 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7501 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7561 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7621 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7681 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7741 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7801 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7861 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7921 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7981 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8041 aagtgccacc tgacgtc
//
