LOCUS       V022386                 7466 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022386
VERSION     V022386
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..2923
                     /label="CD180(NM_005582)"
                     /note="CD180(NM_005582)"
                     /gene="CD180"
     CDS             2963..3028
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3066..3290
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3336..3764
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3778..4107
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4174..4968
                     /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    5142..5263
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5312..5328)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5336..5352
                     /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(5360..5390)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5405..5426
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5714..6299)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6470..7330)
                     /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(7331..7435)
                     /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 atggcgtttg acgtcagctg
      961 cttcttttgg gtggtgctgt tttctgccgg ctgtaaagtc atcacctcct gggatcagat
     1021 gtgcattgag aaagaagcca acaaaacata taactgtgaa aatttaggtc tcagtgaaat
     1081 ccctgacact ctaccaaaca caacagaatt tttggaattc agctttaatt ttttgcctac
     1141 aattcacaat agaaccttca gcagactcat gaatcttacc tttttggatt taactaggtg
     1201 ccagattaac tggatacatg aagacacttt tcaaagccat catcaattaa gcacacttgt
     1261 gttaactgga aatcccctga tattcatggc agaaacatcg cttaatgggc ccaagtcact
     1321 gaagcatctt ttcttaatcc aaacgggaat atccaatctc gagtttattc cagtgcacaa
     1381 tctggaaaac ttggaaagct tgtatcttgg aagcaaccat atttcctcca ttaagttccc
     1441 caaagacttc ccagcacgga atctgaaagt actggatttt cagaataatg ctatacacta
     1501 catctctaga gaagacatga ggtctctgga gcaggccatc aacctaagcc tgaacttcaa
     1561 tggcaataat gttaaaggta ttgagcttgg ggcttttgat tcaacgatct tccaaagttt
     1621 gaactttgga ggaactccaa atttgtctgt tatattcaat ggtctgcaga actctactac
     1681 tcagtctctc tggctgggaa catttgagga cattgatgac gaagatatta gttcagccat
     1741 gctcaaggga ctctgtgaaa tgtctgttga gagcctcaac ctgcaggaac accgcttctc
     1801 tgacatctca tccaccacat ttcagtgctt cacccaactc caagaattgg atctgacagc
     1861 aactcacttg aaagggttac cctctgggat gaagggtctg aacttgctca agaaattagt
     1921 tctcagtgta aatcatttcg atcaattgtg tcaaatcagt gctgccaatt tcccctccct
     1981 tacacacctc tacatcagag gcaacgtgaa gaaacttcac cttggtgttg gctgcttgga
     2041 gaaactagga aaccttcaga cacttgattt aagccataat gacatagagg cttctgactg
     2101 ctgcagtctg caactcaaaa acctgtccca cttgcaaacc ttaaacctga gccacaatga
     2161 gcctcttggt ctccagagtc aggcattcaa agaatgtcct cagctagaac tcctcgattt
     2221 ggcatttacc cgcttacaca ttaatgctcc acaaagtccc ttccaaaacc tccatttcct
     2281 tcaggttctg aatctcactt actgcttcct tgataccagc aatcagcatc ttctagcagg
     2341 cctaccagtt ctccggcatc tcaacttaaa agggaatcac tttcaagatg ggactatcac
     2401 gaagaccaac ctacttcaga ccgtgggcag cttggaggtt ctgattttgt cctcttgtgg
     2461 tctcctctct atagaccagc aagcattcca cagcttggga aaaatgagcc atgtagactt
     2521 aagccacaac agcctgacat gcgacagcat tgattctctt agccatctta agggaatcta
     2581 cctcaatctg gctgccaaca gcattaacat catctcaccc cgtctcctcc ctatcttgtc
     2641 ccagcagagc accattaatt taagtcataa ccccctggac tgcacttgct cgaatattca
     2701 tttcttaaca tggtacaaag aaaacctgca caaacttgaa ggctcggagg agaccacgtg
     2761 tgcaaacccg ccatctctaa ggggagttaa gctatctgat gtcaagcttt cctgtgggat
     2821 tacagccata ggcattttct ttctcatagt atttctatta ttgttggcta ttctgctatt
     2881 ttttgcagtt aaataccttc tcaggtggaa ataccaacac attgaattct gcagatatcc
     2941 agcacagtgg cggccgctcg aggactacaa agaccatgac ggtgattata aagatcatga
     3001 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     3061 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3121 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3181 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3241 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3301 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3361 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3421 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3481 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3541 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3601 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3661 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3721 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3781 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3841 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3961 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4021 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4081 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4141 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4201 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4261 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4321 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4381 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4441 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4501 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4561 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4621 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4681 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4741 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4801 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4861 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4921 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4981 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5041 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5101 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5161 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5221 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5281 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5341 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5401 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5461 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5521 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5581 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5641 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5701 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5761 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5821 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5881 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5941 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6001 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6061 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6121 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6181 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6241 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6361 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6421 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6481 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6541 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6601 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6661 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6721 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6781 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6841 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6901 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6961 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7021 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7081 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7141 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7201 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7261 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7321 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7381 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7441 ttccccgaaa agtgccacct gacgtc
//
