LOCUS       V038298                 8703 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038298
VERSION     V038298
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..3383
                     /label="KPNB1(NM_002265)"
                     /note="KPNB1(NM_002265)"
                     /gene="KPNB1"
     CDS             3390..3455
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3479..4067
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4133..4181
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4383..4811
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4825..5154
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5221..6015
                     /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    6191..6312
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6361..6377)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6385..6401
                     /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(6409..6439)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6454..6475
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6763..7351)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7522..8382)
                     /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(8383..8487)
                     /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 gctgatcacc attctcgaga
      781 agaccgtgtc tcccgatcgg ctggagctgg aagcggcgca gaagttcctg gagcgtgcgg
      841 ccgtggagaa cctgcccact ttccttgtgg aactgtccag agtgctggca aatccaggaa
      901 acagtcaggt tgccagagtt gcagctggtc tacaaatcaa gaactctttg acatctaaag
      961 atccagatat caaggcacaa tatcagcaga ggtggcttgc tattgatgct aatgctcgac
     1021 gagaagtcaa gaactatgtt ttgcagacat tgggtacaga aacttaccgg cctagttctg
     1081 cctcacagtg tgtggctggt attgcttgtg cagagatccc agtaaaccag tggccagaac
     1141 tcattcctca gctggtggcc aatgtcacaa accccaacag cacagagcac atgaaggagt
     1201 cgacattgga agccatcggt tatatttgcc aagatataga cccagagcag ctacaagata
     1261 aatccaatga gattctgact gccataatcc aggggatgag gaaagaagag cctagtaata
     1321 atgtgaagct agctgctacg aatgcactcc tgaactcatt ggagttcacc aaagcaaact
     1381 ttgataaaga gtctgaaagg cactttatta tgcaggtggt ctgtgaagcc acacagtgtc
     1441 cagatacgag ggtacgagtg gctgctttac agaatctggt gaagataatg tccttatatt
     1501 atcagtacat ggagacatat atgggtcctg ctctttttgc aatcacaatc gaagcaatga
     1561 aaagtgacat tgatgaggtg gctttacaag ggatagaatt ctggtccaat gtctgtgatg
     1621 aggaaatgga tttggccatt gaagcttcag aggcagcaga acaaggacgg ccccctgagc
     1681 acaccagcaa gttttatgcg aagggagcac tacagtatct ggttccaatc ctcacacaga
     1741 cactaactaa acaggacgaa aatgatgatg acgatgactg gaacccctgc aaagcagcag
     1801 gggtgtgcct catgcttctg gccacctgct gtgaagatga cattgtccca catgtcctcc
     1861 ccttcattaa agaacacatc aagaacccag attggcggta ccgggatgca gcagtgatgg
     1921 cttttggttg tatcttggaa ggaccagagc ccagtcagct caaaccacta gttatacagg
     1981 ctatgcccac cctaatagaa ttaatgaaag accccagtgt agttgttcga gatacagctg
     2041 catggactgt aggcagaatt tgtgagctgc ttcctgaagc tgccatcaat gatgtctact
     2101 tggctcccct gctacagtgt ctgattgagg gtctcagtgc tgaacccaga gtggcttcaa
     2161 atgtgtgctg ggctttctcc agtctggctg aagctgctta tgaagctgca gacgttgctg
     2221 atgatcagga agaaccagct acttactgct tatcttcttc atttgaactc atagttcaga
     2281 agctcctaga gactacagac agacctgatg gacaccagaa caacctgagg agttctgcat
     2341 atgaatctct gatggaaatt gtgaaaaaca gtgccaagga ttgttatcct gctgtccaga
     2401 aaacgacttt ggtcatcatg gaacgactgc aacaggttct tcagatggag tcacatatcc
     2461 agagcacatc cgatagaatc cagttcaatg accttcagtc tttactctgt gcaactcttc
     2521 agaatgttct tcggaaagtg caacatcaag atgctttgca gatctctgat gtggttatgg
     2581 cctccctgtt aaggatgttc caaagcacag ctgggtctgg gggagtacaa gaggatgccc
     2641 tgatggcagt tagcacactg gtggaagtgt tgggtggtga attcctcaag tacatggagg
     2701 cctttaaacc cttcctgggc attggattaa aaaattatgc tgaataccag gtttgtttgg
     2761 cagctgtggg cttagtggga gacttgtgcc gtgccctgca atccaacatc atacctttct
     2821 gtgacgaggt gatgcagctg cttctggaaa atttggggaa tgagaacgtc cacaggtctg
     2881 tgaagccgca gattctgtca gtgtttggtg atattgccct tgctattgga ggagagttta
     2941 aaaaatactt agaggttgta ttgaatactc ttcagcaggc ctcccaagcc caggtggaca
     3001 agtcagacta tgacatggtg gattatctga atgagctaag ggaaagctgc ttggaagcct
     3061 atactggaat cgtccaggga ttaaaggggg atcaggagaa cgtacacccg gatgtgatgc
     3121 tggtacaacc cagagtagaa tttattctgt ctttcattga ccacattgct ggagatgagg
     3181 atcacacaga tggagtagta gcttgtgctg ctggactaat aggggactta tgtacagcat
     3241 ttgggaagga tgtactgaaa ttagtagaag ctaggccaat gatccatgaa ttgttaactg
     3301 aagggcggag atcgaagact aacaaagcaa aaacccttgc tacatgggca acaaaagaac
     3361 tgaggaaact gaagaaccaa gctggatccg actacaaaga ccatgacggt gattataaag
     3421 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     3481 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     3541 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     3601 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     3661 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     3721 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     3781 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3841 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     3901 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3961 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4021 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     4081 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4141 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4201 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     4261 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     4321 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     4381 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4441 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4501 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     4561 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4621 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4681 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4741 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4801 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4861 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4921 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4981 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5041 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5101 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5161 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5221 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5281 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5341 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5401 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5461 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5521 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5581 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5641 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5701 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5761 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5821 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5881 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5941 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6001 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6061 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6121 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6181 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6241 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6301 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6361 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6421 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     6961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7201 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7321 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7441 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7501 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7621 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7681 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7801 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8461 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8521 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     8581 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     8641 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     8701 agg
//
