LOCUS       V046036                 8910 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046036
VERSION     V046036
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..3590
                     /label="Enpep(NM_007934)"
                     /note="Enpep(NM_007934)"
                     /gene="Enpep"
     CDS             3597..3662
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3686..4274
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4340..4388
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4590..5018
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5032..5361
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5428..6222
                     /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    6398..6519
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6568..6584)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6592..6608
                     /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(6616..6646)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6661..6682
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6970..7558)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7729..8589)
                     /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(8590..8694)
                     /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 ccaccatgaa ctttgcagag gaagagccct
      781 ccaagaaata ctgcattaaa ggcaaacacg tggccatcat ctgtggagta gtggtggcag
      841 ttggattaat agtgggactt tctgtgggtt tgaccaggtc gtgtgagcag gacacaaccc
      901 cagctccttc ccaacctcct ccggaagcca gcactgccct ccctcctcag gaccagaatg
      961 tctgccctga cagtgaagat gaaagcggag aatggaaaaa cttcaggctg cctgacttca
     1021 tcaatccagt tcactacgac ctggaggtga aggccctgat ggaggaagac aggtacacgg
     1081 gaatagtgac catctctgtc aacttgagca aacccactcg tgacctatgg ctccacatca
     1141 gggagaccaa gatcaccaag ctgccggagc taaggaggcc ctctggggag caggtaccaa
     1201 ttcgacggtg cttcgagtat aagaagcagg agtacgtggt gatccaggct gcagaagacc
     1261 ttgcggccac cagtggggac agtgtctatc ggctgaccat ggagtttaaa ggctggctga
     1321 acggttccct tgtgggtttt tacaaaacca cctacatgga ggacgggcaa atcaggagca
     1381 tagctgccac tgaccatgaa ccaacagatg ccaggaagtc cttcccttgt ttcgacgaac
     1441 ccaacaagaa gtcaacttac agtatatcca tcatccaccc aaaagaatac tcagcacttt
     1501 ctaatatgcc agaagagaaa tcagagatgg tggatgacaa ctggaagaaa accacttttg
     1561 tgaagtctgt cccaatgagc acttacctgg tgtgctttgc tgtgcatcgg ttcactgcta
     1621 tagagagaaa atccaggagc ggcaaaccac tcaaggtcta tgtccagccc aatcagaagg
     1681 agacagcaga gtatgcggca aacataaccc aagctgtatt tgattacttc gaagactact
     1741 tcgctatgga gtatgcgctt cctaaactgg ataaaattgc tattccagat tttggcaccg
     1801 gcgccatgga aaattgggga cttgtcactt accgagaaac aaacctgctt tacgaccccc
     1861 tactatcggc ctcatctaac cagcagagag tggccagcgt ggttgcccac gaacttgtac
     1921 accagtggtt tggaaatact gtgaccatgg actggtggga cgacttgtgg ctaaatgaag
     1981 gatttgcttc gttcttcgag ttcttgggag taaaccacgc agaaaaagac tggcagatgc
     2041 tcagtcaggt gctgcttgaa gatgtgttcc ccgtgcaaga ggacgactcc ctgatgtctt
     2101 cacatccagt ggtcgtcacc gtgtccacgc cagctgaaat aacatctgtg tttgatggga
     2161 tatcatacag caagggagct tctattctga gaatgctcca agactggata acaccagaga
     2221 aattccaaaa aggctgtcag atttacttga aaaaattcca gttcgcgaat gcaaaaactt
     2281 ccgacttttg ggattcactg caagaggcaa gcaatctgcc agtgaaagaa gtgatggaca
     2341 cctggactag ccagatgggt tatcctgtgg tcactgtgag tggaaggcag aacatcaccc
     2401 agaaacgctt tctgttggac tccaaagctg atccttcgca gccaccgtca gagctcggtt
     2461 acacatggaa tattccagtc agatgggctg ataatgacaa ctcaaggatc accgtgtaca
     2521 ataggttaga caaaggagga atcactctga atgctaatct tagcggagat gcttttctca
     2581 aaatcaaccc agatcacatt gggttttatc gtgtaaatta tgaaggagga acgtgggatt
     2641 ggatagccga ggctctctcc tcaaaccaca cgagattctc cgctgctgac cggtcaagtt
     2701 ttattgatga tgcttttgct ttggcaagag ctcaacttct gaattataaa atagctctga
     2761 acttgaccat gtatctcaaa tcagaagagg atttcctacc atgggagaga gtcatttcat
     2821 ctgtaagcta catcattagc atgtttgaag atgacagaga gctgtacccc atgatagaga
     2881 cgtactttca aggccaagtg aagcccgttg cagatttgct gggatggcag gataccggaa
     2941 gccacatcac aaagttactc cgggcctcta tcttaggatt cgcatgcaag atgggggaca
     3001 gagaagcctt gggcaatgct tcccagttat ttgactcctg gctgaaaggg agcgcaagta
     3061 ttccggtaaa cctcaggctg cttgtgtacc gctatggaat gcagaactct ggcaatgagg
     3121 cagcgtggaa ctatacccta gagcagtatc agaaaacatc gcttgcccaa gagaaagaaa
     3181 aactgctcta tgggttagct tcagtgaagg atgttaaact cttggcaagg tatctggaaa
     3241 tgctcaaaga ccccaatatt attaaaactc aggatgtatt taccgtcatc cgctacatct
     3301 cctacaacag ttatgggaag acaatggcct ggaattggat acaactcaac tgggactatc
     3361 tggtcagcag atttacaatc aatgacagat accttggccg gatcgtcacc atagctgagc
     3421 ccttcaacac tgaactgcag ctctggcaga tgcagagctt ttttgcaaaa tatccaaatg
     3481 ctggcgctgg agcaaaacct agagagcaag tgctggagac ggtgaagaac aacatcgagt
     3541 ggctaaacgt gaacagacag tccatcagag agtggttcgc tagcctgccg ggatccgact
     3601 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     3661 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     3721 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3781 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3841 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3901 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3961 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4021 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4081 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4141 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4201 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4261 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     4321 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     4381 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     4441 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     4501 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     4561 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4621 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4681 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4741 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4801 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4861 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4921 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     4981 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     5041 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5101 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5161 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5221 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5281 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     5341 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     5401 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5461 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5521 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5581 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5641 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5701 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5761 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5821 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5881 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5941 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6001 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6061 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6121 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6181 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6241 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6301 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6361 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     6421 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6481 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6541 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6721 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6781 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6841 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6901 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6961 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7021 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7081 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7141 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     7201 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7261 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7321 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7381 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7441 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7501 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7561 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7621 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7681 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7741 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7801 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7861 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7921 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7981 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8041 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8101 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8161 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8221 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8281 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8341 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8401 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8461 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8521 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8581 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8641 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8701 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     8761 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     8821 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     8881 accgacaatt gcatgaagaa tctgcttagg
//
