pRK5-HA-Ubiquitin-K48人源基因质粒

质粒图谱

质粒序列

LOCUS       Exported                4996 bp ds-DNA     circular SYN 27-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 27, 2017 from Plasmid
FEATURES             Location/Qualifiers
     source          1..4996
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      963..972
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             975..1001
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1032..1259
                     /codon_start=1
                     /product="human ubiquitin variant containing Lys-48, with 
                     all other lysines mutated to arginine"
                     /label=ubiquitin-K48
                     /translation="MQIFVRTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQRESTLHLVLRLRGG"
     regulatory      1296..1305
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    1309..1443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1512..1869
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1720..1855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      2118..2573
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2855..2959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2960..3820
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3991..4579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc aaggccattc
      961 gcgccaccat gggctacccc tatgatgtgc ctgactacgc agatctcaat ggtggtggtg
     1021 gtgggtcgac catgcagatc ttcgtcagaa cgttaaccgg tagaaccata actctagaag
     1081 ttgaaccatc cgataccatc gaaaacgtta gagctagaat tcaagacaga gaaggcattc
     1141 cacctgatca acaaagattg atctttgccg gtaagcagct cgaggacggt agaacgctgt
     1201 ctgattacaa cattcagaga gagtccaccc tgcacctggt cctccgtctc agaggtggtt
     1261 gagcggccgc taagtaagta aggatcccca gcttggccgc catggcccaa cttgtttatt
     1321 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1381 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     1441 atcgggaatt aattcggcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     1501 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     1561 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1621 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1681 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     1741 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     1801 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     1861 ttttgcaaaa agctgttaac agcttggcac tggccgtcgt tttacaacgt cgtgactggg
     1921 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     1981 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2041 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2101 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2161 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2221 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2281 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2341 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2401 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     2461 ctattctttt aatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2521 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     2581 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2641 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2701 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2761 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2821 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2881 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2941 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3001 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3061 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3121 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3181 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3241 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3301 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3361 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3421 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3481 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3541 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3601 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3661 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3721 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3781 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3841 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3901 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3961 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4021 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4081 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4141 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4201 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4261 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4321 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4381 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4441 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4501 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4561 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4621 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4681 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4741 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4801 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     4861 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4921 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     4981 catgattacg aattaa