LOCUS       V046180                 7215 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046180
VERSION     V046180
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..1895
                     /label="Nup43(NM_145706)"
                     /note="Nup43(NM_145706)"
                     /gene="Nup43"
     CDS             1902..1967
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1991..2579
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2645..2693
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2895..3323
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3337..3666
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3733..4527
                     /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    4703..4824
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4873..4889)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4897..4913
                     /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(4921..4951)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4966..4987
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5275..5863)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6034..6894)
                     /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(6895..6999)
                     /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 ccaccatgga ggaaatttat gctaagttcg
      781 tgtctcagaa aattagcaaa acccgctggc ggccagtgcc ttccgggagc ctgcagacca
      841 cagagacctt tgccacgggc tcttgggaca atgaggaaaa ttgcgtttca ctgtggtcta
      901 ttggagattt tggaaacttg gattctgacg gagggtttga aggagaccat cagttactgt
      961 gcgatattag acatcatggg gatgtcatgg atttacagtt ttttgaccag gaaagaattg
     1021 tagctgcttc gtcaacaggc tgtgtaacag tttttcttca ccatccaaat aaccagaccc
     1081 tgtccgtcaa tcagcagtgg cctgcagctc attaccatac aggtcccagc agtccctcct
     1141 atagcagtgc accatgtaca ggaattgtgt gtgacaaccc agaaattgtt acagttggag
     1201 aggatggccg aataaatctc ttcagagttg atcacaagga ggctgtcaga actatagata
     1261 atgcagacag cagtacactc catgccgtaa ccttccttcg aactcctgag attgttacag
     1321 taaattcaat tggacagtta aaaatatggg atttcagaca acaaggaagt gaaccctgtc
     1381 agatactatc actgactggt gaccgagtgc cactccactg tgttgacaga catccagacc
     1441 agcagcatgt tgtagctacc ggtggccagg atggaatgtt gagtatttgg gatgtcagac
     1501 aaggtaccat gcctgtgtca ttactgaagg ctcatgaggc tgaaatgtgg gaggttcact
     1561 tccacccgtc caacccagat cacctgttca cttgttctga agatggatct ctctggcatt
     1621 gggatgcctc cacagatgca cctgaaaagt cttctctctt tcaccaagga ggaagaagta
     1681 gtactttcct gtcacatagc ttaagtaacc aggccggtgt tcaccagtct ctagtaagtt
     1741 cctggctcag cacagaccct gccaaagacc gaattgaaat cacaagcttg cttcccagca
     1801 ggactctgtc tgtaaacagt ttggatgttt taggtccctg tcttgtttgt ggaactgatg
     1861 cagaagccat ttatgttacc agacaactat tttcaggatc cgactacaaa gaccatgacg
     1921 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     1981 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2041 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2101 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2161 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2221 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2281 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2341 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     2401 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2461 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2521 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     2581 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     2641 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     2701 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     2761 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     2821 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     2881 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2941 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3001 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3061 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3121 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3181 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3241 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     3301 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3361 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3421 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3481 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3541 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3601 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3661 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3721 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3781 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3841 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3901 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3961 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4021 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4081 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4141 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4201 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4261 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4321 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4381 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4441 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4501 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     4561 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4621 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4681 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4741 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4801 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4861 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4921 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4981 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5041 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5101 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5461 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5701 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7021 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     7081 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7141 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7201 aagaatctgc ttagg
//
