LOCUS       V033445                 7950 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033445
VERSION     V033445
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             756..2630
                     /label="IRAK2(NM_001570)"
                     /note="IRAK2(NM_001570)"
                     /gene="IRAK2"
     CDS             2637..2702
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2726..3314
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3380..3428
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3630..4058
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4072..4401
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4468..5262
                     /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    5438..5559
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5608..5624)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5632..5648
                     /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(5656..5686)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5701..5722
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6010..6598)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6769..7629)
                     /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(7630..7734)
                     /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 ctcgaattcg ccaccatggc ctgctacatc taccagctgc
      781 cctcctgggt gctggacgac ctgtgccgca acatggacgc gctcagcgag tgggactgga
      841 tggagttcgc ctcctacgtg atcacagacc tgacccagct gcggaagatc aagtccatgg
      901 agcgggtgca gggtgtgagc atcacgcggg agctgctgtg gtggtggggc atgcggcagg
      961 ccaccgtcca gcaacttgtg gacctcctgt gccgcctgga gctctaccgg gctgcccaga
     1021 tcatcctgaa ctggaaaccg gctcctgaaa tcaggtgtcc cattccagcc ttccctgact
     1081 ctgtgaagcc agaaaagcct ttggcagctt ctgtaagaaa ggctgaggat gaacaggaag
     1141 aggggcagcc tgtgaggatg gccacctttc caggcccagg gtcctctcca gccagagccc
     1201 accagccggc ctttctccag cctcctgaag aagatgcccc tcattccttg agaagcgacc
     1261 tccccacttc gtctgattca aaggacttca gcacctccat tcctaagcag gaaaaacttt
     1321 tgagcttggc tggagacagc cttttctgga gtgaggcaga cgtggtccag gcaaccgatg
     1381 acttcaatca aaaccgcaaa atcagccagg ggacctttgc tgacgtctac agagggcaca
     1441 ggcacgggaa gccattcgtc ttcaagaagc tcagagagac agcctgttca agtccaggat
     1501 caatcgaaag attcttccag gcagagttgc agatttgtct tagatgctgc caccccaatg
     1561 tcttacctgt gctgggcttc tgtgctgcaa gacagtttca cagcttcatc tacccctaca
     1621 tggcaaatgg ttccctacag gacagactgc agggtcaggg tggctcggac cccctcccct
     1681 ggccccagcg tgtcagcatc tgctcagggc tgctctgtgc cgtcgagtac ctgcatggtc
     1741 tggagatcat ccacagcaac gtcaagagct ctaatgtctt gctggaccaa aatctcaccc
     1801 ccaaacttgc tcacccaatg gctcatctgt gtcctgtcaa caaaaggtca aaatacacca
     1861 tgatgaagac tcacctgctc cggacgtcag ccgcgtatct gccagaggat ttcatccggg
     1921 tggggcagct gacaaagcga gtggacatct tcagctgtgg aatagtgttg gccgaggtcc
     1981 tcacgggcat ccctgcaatg gataacaacc gaagcccggt ttacctgaag gacttactcc
     2041 tcagtgatat tccaagcagc accgcctcgc tctgctccag gaagacgggc gtggagaacg
     2101 tgatggcaaa ggagatctgc cagaagtacc tggagaaggg cgcagggagg cttccggagg
     2161 actgcgccga ggccctggcc acggctgcct gcctgtgcct gcggaggcgt aacaccagcc
     2221 tgcaggaggt gtgtggctct gtggctgctg tggaagagcg gctccgaggt cgggagacgt
     2281 tgctcccttg gagtgggctt tctgagggta caggctcttc ttccaacacc ccagaggaaa
     2341 cagacgacgt tgacaattcc agccttgatg cctcctcctc catgagtgtg gcaccctggg
     2401 caggggctgc caccccactt ctccccacag agaatgggga aggaaggctg cgggtcatcg
     2461 tgggaaggga ggctgactcc tcctctgagg cctgtgttgg cctggagcct ccccaggatg
     2521 ttacagaaac ttcgtggcaa attgagatca atgaggccaa aaggaaactg atggagaata
     2581 ttctgctcta caaagaggaa aaagtggaca gcattgagct ctttggcccc ggatccgact
     2641 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     2701 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     2761 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2821 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2881 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2941 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3001 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3061 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3121 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3181 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3241 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3301 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     3361 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     3421 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3481 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3541 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     3601 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3661 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3721 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3781 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3841 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3901 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3961 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     4021 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4081 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4201 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4261 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4321 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4381 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4441 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4501 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4561 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4621 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4681 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4741 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4801 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4861 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4921 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4981 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5041 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5101 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5161 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5221 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5281 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5341 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5401 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5461 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5521 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5581 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6181 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     6241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6421 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6541 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6601 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6661 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6721 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6781 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6841 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6901 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6961 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7021 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7081 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7141 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7201 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7261 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7321 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7381 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7441 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7501 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7561 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7621 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7681 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7741 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     7801 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     7861 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     7921 accgacaatt gcatgaagaa tctgcttagg
//
