LOCUS       V022814                 7276 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022814
VERSION     V022814
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2829
                     /label="MAU2(NM_015329)"
                     /note="MAU2(NM_015329)"
                     /gene="MAU2"
     polyA_signal    2876..3100
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3146..3574
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3588..3917
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3984..4778
                     /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    4952..5073
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5122..5138)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5146..5162
                     /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(5170..5200)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5215..5236
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5524..6109)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6280..7140)
                     /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(7141..7245)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggcggctc aggcggcggc agcggcccag
     1021 gcggcggcgg cccaggctgc gcaggccgag gcggccgact cgtggtacct ggcgcttctg
     1081 ggcttcgctg agcacttccg cacttccagc ccgcccaaaa tccgcctgtg cgtgcactgc
     1141 ctgcaggccg tgttcccctt caagccgccg cagcgcatcg aggcccgtac acacctgcag
     1201 ctgggctccg ttctctatca ccacaccaag aacagcgagc aggcgcgcag ccacctggag
     1261 aaggcgtggt tgatatcaca gcaaatcccg cagttcgaag atgttaaatt tgaagcagca
     1321 agtctgttgt ctgaattgta ctgtcaagag aattccgttg atgcagcaaa gccgctgctg
     1381 cggaaggcga tccagatctc acagcagacc ccatattggc actgccgcct gctcttccag
     1441 ctcgctcaac tgcacacgct tgagaaggac ctggtgtcgg cctgtgacct cctgggtgta
     1501 ggggccgagt acgcccgggt ggtgggatct gaatacacac gggcgctgtt cctcctcagc
     1561 aaggggatgc tgctgctgat ggagcgaaag ctgcaggagg tgcacccgct gctgaccctc
     1621 tgcgggcaga tcgtggagaa ctggcagggg aaccccatcc agaaggagtc gctgcgtgtc
     1681 ttcttcctgg tgctccaggt cacccactat ctggatgccg ggcaggtgaa gagcgtgaag
     1741 ccgtgtctga agcagctgca gcagtgcatc cagaccatct ccacactgca cgatgatgag
     1801 atcctgccca gcaaccccgc tgacctcttc cactggctgc ccaaggagca catgtgtgtg
     1861 cttgtctacc tggtgactgt gatgcactcc atgcaggccg gctacctgga gaaggcgcag
     1921 aagtacacgg acaaggccct catgcagctg gagaagctca agatgctgga ctgcagcccc
     1981 atcctgtcat ccttccaagt gatcctgctg gagcacatca tcatgtgccg ccttgtcacg
     2041 ggtcacaagg ccacggcgct gcaggagatc tcccaggtct gccagctgtg ccagcagtcc
     2101 ccccggctct tctccaacca tgcagcacag ctgcacacat tgctgggcct gtactgtgtc
     2161 tctgtcaact gcatggacaa cgcggaagcc cagttcacca cggccctgcg gctcaccaac
     2221 caccaggagc tgtgggcctt catcgtcacc aacctggcga gtgtgtatat acgggaagga
     2281 aatagacacc aagaggtact ctacagtctg ctggagagga tcaacccgga ccacagcttc
     2341 cctgtcagct cgcactgcct ccgagcagcc gccttctatg tgcgtgggct cttctccttc
     2401 ttccagggac gctacaacga ggccaagcga tttctgcggg aaactctgaa gatgtccaat
     2461 gctgaggacc tgaaccggct cacagcctgc tccctcgtgc ttctgggcca catcttctat
     2521 gtgctgggaa accacaggga gagtaacaac atggtggtgc ctgccatgca gctcgccagc
     2581 aagatcccgg acatgtcggt acagctgtgg tcgtcagcac tgctgagaga cctgaataaa
     2641 gcctgtggga acgccatgga tgcccatgaa gccgcccaga tgcaccagaa cttctcgcag
     2701 cagctgctcc aggaccacat tgaggcctgc agcctccccg aacacaacct catcacgtgg
     2761 acagacggtc caccccccgt gcagttccaa gctcagaatg gacccaacac cagcctggcc
     2821 agcctcctgt gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2881 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2941 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3001 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3061 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3121 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3181 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3241 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3301 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3361 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3421 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3481 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3541 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3661 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3721 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3781 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3841 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3901 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3961 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4021 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4081 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4141 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4201 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4261 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4321 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4381 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4441 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4501 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4561 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4621 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4681 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4741 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4801 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4861 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4921 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4981 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5041 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5101 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5161 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5221 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5281 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5341 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5401 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5461 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5521 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5581 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5641 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5701 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5761 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5821 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5881 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5941 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6001 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6061 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6121 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6181 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6241 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6301 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6361 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6421 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6481 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6541 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6601 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6661 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6721 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6781 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6841 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6901 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6961 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7021 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7081 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7141 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7201 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7261 agtgccacct gacgtc
//
