LOCUS       V046223                10461 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046223
VERSION     V046223
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..5141
                     /label="Nup153(NM_175749)"
                     /note="Nup153(NM_175749)"
                     /gene="Nup153"
     CDS             5148..5213
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5237..5825
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5891..5939
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6141..6569
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6583..6912
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6979..7773
                     /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    7949..8070
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8119..8135)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8143..8159
                     /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(8167..8197)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8212..8233
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8521..9109)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9280..10140)
                     /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(10141..10245)
                     /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 ctcggaagcc ggaggcatag
      781 gagggggcgg cggcggcggc aagatccgga ctcgtcgttg ccaccagggg ccggtgaagc
      841 cttaccaaca ggggcgaccc caacaccagg gcattcttag cagggttaca gaatctgtta
      901 agaatattgt accagggtgg ctacaaagat acttcaacaa gagtgaaaat gcttgcagct
      961 gttcaccaga tgcagatgaa gttccaccct ggccagaaaa tagagaagat gagcatgcga
     1021 tttatgcgga tgagaacaca aacactgatg atgggagaat cacccctgac cctgcaggca
     1081 gtaacacaga agaaccatca acaaccagta ctgcttcaaa ttatccagat gtgttaacaa
     1141 gaccttctct tcatcggagt catctgaatt tctctgtgtt ggaatcccct gctttgcact
     1201 gtcagccgtc cacatcctct gcgttcccaa tcggcagttc tggattttcc cttgtgaagg
     1261 agattaaaga ttctacctct cagcatgatg atgataacat ctcaactacc agtgggttct
     1321 cttcaagagc ttctgagaaa gatatagctg tttcaaagaa cacttccctg ccacctttgt
     1381 ggtcccctga agcagaacgt tctcattcac tctcacagca tactgccatc agctccaaaa
     1441 aaccagcatt caacttgtct gcctttggaa cactttccac ctcacttggg aattcttcaa
     1501 ttcttaaaac aagtcagctt ggagactctc ctttttatcc tggcaaaaca acatatggag
     1561 gggcagctgc tgctgtaaga cagaataaag tacgaagtac accttaccag gctccagtta
     1621 gaagacagat gaaagctaag cagctcaatg cacagtctta tggtgtgacc agctcaacag
     1681 cacggcggat actgcagtca ttagaaaaga tgtcgagccc tcttgcggat gcaaaaagaa
     1741 ttccatctgc agtttcttct cctctgaatt ctcctcttga taggagtggg atagataaca
     1801 cagtttttca ggcaaaaaaa gaaaaggtgg actctcagta tccccctgtt cagagactaa
     1861 tgaccccaaa gccagtttcc atagcaacaa atagaactgt ctattttaaa ccatctctga
     1921 ccccatctgg tgacttaagg aagactaatc aaaggataga taaaaagaac agtactgtag
     1981 atgaaaaaag tatctcaaga caaaatagag agcaagaaag tggcttttca taccccaatt
     2041 tcagtattcc tgcagccaat ggtttatctt ctggagtagg tggtggaggt ggcaagatga
     2101 gaagagagag aacacatttt gttgctccta aacctccaga gaatgaggaa gtggaagcac
     2161 cattgttacc gcaaatctct ctaccaatca gcagttcttc actgcccacc ttcagtttta
     2221 gttcccctgt gacctcggct tctccttcac cagtcagctc ttctcagcca ttaccaaaca
     2281 aggtacagat gacctctctg ggcagcacgg gcagtcctgt gttcacattc tcatctccca
     2341 ttgtaaaatc cacgcaggca gctgtgctcc ctccagcatc tattggattt acatttagtg
     2401 tgcctcttgc aaaaactgaa ttttctggct ctaatagttc ttcagaaaca gttttaagta
     2461 gttcagccca agatatcact gcagtgaata gttcaagcta taagaagcga agtgctcctt
     2521 gtgaggaccc ctttacacct gcaaagatcc tgagagaagg aagtgttcta gacattctga
     2581 aaacacctgg tttcgcatca ccaaaggttg attctcctgc tcttcaaccc actaccacaa
     2641 gctcaatagt ttatacaaga ccagcaataa gtactttttc ttctagtgga attgagtatg
     2701 gagaaagttt aaaagctgga tcatcatggc agtgtgacac atgtctcctc cagaacaaag
     2761 tcacagacaa taagtgtata gcctgccaag cagcaaaact gcccctgaaa gaaactgcta
     2821 aacagactgg gactgggaca ccaagcaaaa gtgacaaacc agcctctaca tcagggacag
     2881 gctttggaga taaatttaaa ccagcaatag gaacttggga ttgtgatacc tgtttagtgc
     2941 aaaataagcc cgaagcagta aaatgtgtag cctgtgagac acctaagcct ggcactggtg
     3001 tgaagcgagc ccttacattg acagtggctt cagaaagtcc tgtgactgct tcctcttcta
     3061 ccactgtgac aactggtaca ctaggatttg gagataaatt caaaaggcct gtgggctctt
     3121 gggagtgtcc agtctgctgt gtttgtaata aggcagaaga caacagatgt gtttcctgta
     3181 catctgagaa accaggtttg gtatctgcct caagcagcag ccctgcccct gtctctctgt
     3241 cttctggagg ctgtttagga ttggacaagt ttaaaaagcc cgagggaagc tgggactgtg
     3301 aagtgtgcct agtgcaaaat aaagcagact ctgccaagtg catagcttgt gaaagtgcaa
     3361 agccaggcac aaagtctgag tttaaaggct tcgggacatc ttcatcttta aatccagcac
     3421 cctcagcctt caaatttggt attccatcgt cctcttctgg gctttctcaa actttgacaa
     3481 gcactggaaa ttttaaattt ggagatcagg gaggattcaa attaggcact tcatctgact
     3541 ctgggtctac aaacaccatg aatacaaact ttaaattttc taaaccaact ggagacttta
     3601 aatttggagt gttatctgat tctaaacctg aagaagttaa aaacgacaac aagaatgata
     3661 attttcagtt tggatcttct tctggcttaa ccaacccagc ttcttcagct ccgtttcagt
     3721 ttggggtatc tactcttggg caacaagaaa agaaagagga gctgcctaaa tcttcacctg
     3781 caggctttag ctttggtgca ggtgttaata acccccctaa tgctgctatt gacaccacag
     3841 cgacctcaga gaacaagagc ggcttcaact ttggaaccct agacacaaag agtgtctcag
     3901 tgactccttt cacatacaag acaacagaag caaagaaaga agacgcacct gctaccaagg
     3961 gtgggttcac atttggtaaa gtgggctcct cttctttgcc atcttcctca atgtttgttt
     4021 tgggaaggac agaggagaaa cagcaagaac ctgttacttc tacttctctg gtgtttggga
     4081 agaaagccga cagtgaggag ccaaagtgtc agccagtatt ttcctttggg aattcagagc
     4141 aaaccaaaga tgagagctcc aagccgacat tcagcttcag tgtggcaaaa ccatctggga
     4201 aggagtctga gcagctagcc aaggctactt ttgcatttgg aaatcaaacc aatacaacaa
     4261 ctgatcaagg tgcagcaaag ccagttttta gcttcttgaa cagcagttcc tctagttcaa
     4321 gtgcaccagc cacttcatct agtggtggca tatttggtag ttccacctct tcctccaacc
     4381 cacctgtggc agcctttgtg tttggacagg ccagcaatcc tgtcagcagc tctgccttcg
     4441 gtaacgctgc tgaatccagt acatctcagt ctttgctatt tcctcaagag agcgagccag
     4501 ccaccacgtc cagcactgcc ccagctgcct ctccctttgt atttgggaca ggagccagca
     4561 gtaatagtgt gtcttctggg ttcacttttg gggctacaac cacgtcaagc tcttcaggat
     4621 cctcctttgt atttggcact ggacattcag caccatctgc cagtccagca tttggtgcta
     4681 accagactcc aacatttgga caaagtcaag gtgccagcca gcctaacccg ccaagctttg
     4741 gctctatctc gtcttcaaca gcattatttt ctgctggttc tcagcctgta ccaccaccta
     4801 tatttgggac agtgtcaagc agcagccaac ctcctgtgtt tggacagcag cctagtcagt
     4861 ctgcatttgg ctctgggaca gctaatgcca gttcggtttt ccaatttggt agcagcacta
     4921 cgaatttcaa cttcacaaac aataatccat caggagtgtt cacatttggt gcaagtccca
     4981 gcacgcctgc agcctcagcc cagccctcag gttcaggggt cttttcattc agccagtctc
     5041 cagcatcatt tacagtgggg tcaaatggga aaaatatgtt ctcttcttcc ggaacttcag
     5101 tttctggtcg caagataaag actgctgtta gacgcaagaa aggatccgac tacaaagacc
     5161 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     5221 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5281 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5341 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5401 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5461 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5521 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5581 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     5641 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5701 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5761 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5821 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5881 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     5941 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6001 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6061 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     6121 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6181 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6241 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     6301 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6361 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6421 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6481 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     6541 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     6601 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6661 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6721 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6781 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6841 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     6901 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     6961 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     7021 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     7081 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     7141 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     7201 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     7261 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     7321 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     7381 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     7441 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     7501 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     7561 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     7621 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7681 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7741 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     7801 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     7861 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     7921 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     7981 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     8041 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     8101 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     8161 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     8221 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     8281 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     8341 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     8401 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     8461 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     8521 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     8581 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8641 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8701 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8761 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8821 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8881 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8941 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     9001 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     9061 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     9121 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     9181 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     9241 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     9301 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     9361 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     9421 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     9481 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     9541 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     9601 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     9661 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     9721 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9781 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9841 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9901 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9961 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    10021 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    10081 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    10141 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    10201 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    10261 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
    10321 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
    10381 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
    10441 tgcatgaaga atctgcttag g
//
