pRPa series

Members of this plasmid series integrate at the tagged rDNA spacer locus in the 2T1 ‘landing pad’ cell line following AscI linearisation at the HYG and RRNA spacer targeting sequences.

Derived from known sequence and junction sequencing, February 2008

Landmarks Available expression cassettes
0001-0600: (600) RRNA targeting fragment AscI Sequence page Expression cassette Region replaced
0601-3124: (2525) Vector backbone (pUC/pGEM) Stem-loop RNAi MCS1/2 Acc65I – Bsp120I
3125-3186: (62) Remnant of T. brucei actin mRNA processing 3’ MCS3/4
3187-4207: (1021) Hygromycin B phosphotransferase ORF ΔSTOP AscI
4208-4465: (258) T. brucei aldolase mRNA processing 5’ N-term eGFP GFPx HindIII – BamHI
4449-4682: (234) EP procyclin promoter C-term eGFP xGFP
4688-4947: (260) RRNA promoter N-term MYC MYCx
4951-4963: (13) Tet operator N/C-term 6MYC x6MYCx
4970-5064: (95) T. brucei procyclin mRNA processing 5′
5062-7596: (2535) Expression Cassette (GFPx = 2,535 bp) Constitutive GFPx NheI – Acc65I
7597-8334: (738) T. brucei aldolase mRNA processing 3’
8335-8736: (402) T7 terminator x2 fragment
    1  GAATTCGATG CGAGGCGAAT CGCTCAGTAG TGTTAGTGCA GGCATTGTTT
   51  CTCTTTTCCA TTTCGGCTCG CTCTCTGTGC CAGAAAGGCT TTTGGAGTCT
  101  TGGTTACTCG TGGGACTAGG GCAAGCGTCG CTTCCCTAAT AATTGTTGCT
  151  AACGTTTCAC CGTGCGTGTG TGGATATCTT CCAAGTTGTC ATTTTCTCTA
  201  CCCCTCTCTT TTCCGGCGCG GTCTAACTTC CTCCAACTAG CGCATTTCCA
  251  TGATAACGGG GTCCACATAG TCCGTTGTAG TGAAATTGGC ATGCCATCAG
  301  GGCCAAGGCA TACATCGCGG CCGCGATAAA AATGATACGA TGATCTCCGT
  351  GTGGAGCAGA CCGTGGCGAC CAGTTTGTAC CATGGAAGTT CTTCGATAGA
  401  GTTATATCGA GTGCGGAATT GATGCTACGG TCACAGTGTC GTATCGTGTG
  451  GTCTCCCATG TGTTTTTGAC GGTGTACCAA TTATCTTTGC ACTAATGGGC
  501  TATGGAATCT TTCCTTTTAA TCAGGGGGAC ATGTTTATCC CACGCCAGTA
  551  GTCTGGAATT AGAATACATA CCAACTATAT GGCGCGCCGA GCTCGAATTC
  601  TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG
  651  CGCTCTTCCG CTTCCTCGCT CACTGACTCG CTGCGCTCGG TCGTTCGGCT
  701  GCGGCGAGCG GTATCAGCTC ACTCAAAGGC GGTAATACGG TTATCCACAG
  751  AATCAGGGGA TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG
  801  GCCAGGAACC GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG
  851  CCCCCCTGAC GAGCATCACA AAAATCGACG CTCAAGTCAG AGGTGGCGAA
  901  ACCCGACAGG ACTATAAAGA TACCAGGCGT TTCCCCCTGG AAGCTCCCTC
  951  GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT ACCGGATACC TGTCCGCCTT
 1001  TCTCCCTTCG GGAAGCGTGG CGCTTTCTCA TAGCTCACGC TGTAGGTATC
 1051  TCAGTTCGGT GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC
 1101  CCCGTTCAGC CCGACCGCTG CGCCTTATCC GGTAACTATC GTCTTGAGTC
 1151  CAACCCGGTA AGACACGACT TATCGCCACT GGCAGCAGCC ACTGGTAACA
 1201  GGATTAGCAG AGCGAGGTAT GTAGGCGGTG CTACAGAGTT CTTGAAGTGG
 1251  TGGCCTAACT ACGGCTACAC TAGAAGAACA GTATTTGGTA TCTGCGCTCT
 1301  GCTGAAGCCA GTTACCTTCG GAAAAAGAGT TGGTAGCTCT TGATCCGGCA
 1351  AACAAACCAC CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT
 1401  ACGCGCAGAA AAAAAGGATC TCAAGAAGAT CCTTTGATCT TTTCTACGGG
 1451  GTCTGACGCT CAGTGGAACG AAAACTCACG TTAAGGGATT TTGGTCATGA
 1501  GATTATCAAA AAGGATCTTC ACCTAGATCC TTTTAAATTA AAAATGAAGT
 1551  TTTAAATCAA TCTAAAGTAT ATATGAGTAA ACTTGGTCTG ACAGTTACCA
 1601  ATGCTTAATC AGTGAGGCAC CTATCTCAGC GATCTGTCTA TTTCGTTCAT
 1651  CCATAGTTGC CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC
 1701  TTACCATCTG GCCCCAGTGC TGCAATGATA CCGCGAGACC CACGCTCACC
 1751  GGCTCCAGAT TTATCAGCAA TAAACCAGCC AGCCGGAAGG GCCGAGCGCA
 1801  GAAGTGGTCC TGCAACTTTA TCCGCCTCCA TCCAGTCTAT TAATTGTTGC
 1851  CGGGAAGCTA GAGTAAGTAG TTCGCCAGTT AATAGTTTGC GCAACGTTGT
 1901  TGCCATTGCT ACAGGCATCG TGGTGTCACG CTCGTCGTTT GGTATGGCTT
 1951  CATTCAGCTC CGGTTCCCAA CGATCAAGGC GAGTTACATG ATCCCCCATG
 2001  TTGTGCAAAA AAGCGGTTAG CTCCTTCGGT CCTCCGATCG TTGTCAGAAG
 2051  TAAGTTGGCC GCAGTGTTAT CACTCATGGT TATGGCAGCA CTGCATAATT
 2101  CTCTTACTGT CATGCCATCC GTAAGATGCT TTTCTGTGAC TGGTGAGTAC
 2151  TCAACCAAGT CATTCTGAGA ATAGTGTATG CGGCGACCGA GTTGCTCTTG
 2201  CCCGGCGTCA ATACGGGATA ATACCGCGCC ACATAGCAGA ACTTTAAAAG
 2251  TGCTCATCAT TGGAAAACGT TCTTCGGGGC GAAAACTCTC AAGGATCTTA
 2301  CCGCTGTTGA GATCCAGTTC GATGTAACCC ACTCGTGCAC CCAACTGATC
 2351  TTCAGCATCT TTTACTTTCA CCAGCGTTTC TGGGTGAGCA AAAACAGGAA
 2401  GGCAAAATGC CGCAAAAAAG GGAATAAGGG CGACACGGAA ATGTTGAATA
 2451  CTCATACTCT TCCTTTTTCA ATATTATTGA AGCATTTATC AGGGTTATTG
 2501  TCTCATGAGC GGATACATAT TTGAATGTAT TTAGAAAAAT AAACAAATAG
 2551  GGGTTCCGCG CACATTTCCC CGAAAAGTGC CACCTGACGT CTAAGAAACC
 2601  ATTATTATCA TGACATTAAC CTATAAAAAT AGGCGTATCA CGAGGCCCTT
 2651  TCGTCTCGCG CGTTTCGGTG ATGACGGTGA AAACCTCTGA CACATGCAGC
 2701  TCCCGGAGAC GGTCACAGCT TGTCTGTAAG CGGATGCCGG GAGCAGACAA
 2751  GCCCGTCAGG GCGCGTCAGC GGGTGTTGGC GGGTGTCGGG GCTGGCTTAA
 2801  CTATGCGGCA TCAGAGCAGA TTGTACTGAG AGTGCACCAT TCGACGCTCT
 2851  CCCTTATGCG ACTCCTGCAT TAGGAAGCAG CCCAGTAGTA GGTTGAGGCC
 2901  GTTGAGCACC GCCGCCGCAA GGAATGGTGC AAGGAGATGG CGCCCAACAG
 2951  TCCCCCGGCC ACGGGGCCTG CCACCATACC CACGCCGAAA CAAGCGCTCA
 3001  TGAGCCCGAA GTGGCGAGCC CGATCTTCCC CATCGGTGAT GTCGGCGATA
 3051  TAGGCGCCAG CAACCGCACC TGTGGCGCCG GTGATGCCGG CCACGATGCG
 3101  TCCGGCGTAG AGGATCTGGC TAGGAATACT GCATAGATAA CAAACGCATC
 3151  AAACACAACT GGTGGCACTA TTTCAATCAT GTCGACGGCG CGCCCTCGGA
 3201  CGAGTGCTGG GGCGTCGGTT TCCACTATCG GCGAGTACTT CTACACAGCC
 3251  ATCGGTCCAG ACGGCCGCGC TTCTGCGGGC GATTTGTGTA CGCCCGACAG
 3301  TCCCGGCTCC GGATCGGACG ATTGCGTCGC ATCGACCCTG CGCCCAAGCT
 3351  GCATCATCGA AATTGCCGTC AACCAAGCTC TGATAGAGTT GGTCAAGACC
 3401  AATGCGGAGC ATATACGCCC GGAGCCGCGG CGATCCTGCA AGCTCCGGAT
 3451  GCCTCCGCTC GAAGTAGCGC GTCTGCTGCT CCATACAAGC CAACCACGGC
 3501  CTCCAGAAGA AGATGTTGGC GACCTCGTAT TGGGAATCCC CGAACATCGC
 3551  CTCGCTCCAG TCAATGACCG CTGTTATGCG GCCATTGTCC GTCAGGACAT
 3601  TGTTGGAGCC GAAATCCGCG TGCACGAGGT GCCGGACTTC GGGGCAGTCC
 3651  TCGGCCCAAA GCATCAGCTC ATCGAGAGCC TGCGCGACGG ACGCACTGAC
 3701  GGTGTCGTCC ATCACAGTTT GCCAGTGATA CACATGGGGA TCAGCAATCG
 3751  CGCATATGAA ATCACGCCAT GTAGTGTATT GACCGATTCC TTGCGGTCCG
 3801  AATGGGCCGA ACCCGCTCGT CTGGCTAAGA TCGGCCGCAG CGATCGCATC
 3851  CATGGCCTCC GCGACCGGCT GCAGAACAGC GGGCAGTTCG GTTTCAGGCA
 3901  GGTCTTGCGT GACACCCTGT GCACGGCGGG AGATGCAATA GGTCAGGCTC
 3951  TCGCTGAATT CCCCAATGTC AAGCACTTCC GGAATCGGGA GCGCGGCCGA
 4001  TGCAAAGTGC CGATAAACAT AACGATCTTT GTAGAAACCA TCGGCGCAGC
 4051  TATTTACCCG CAGGACATAT CCACGCCCTC CTACATCGAA GCTGAAAGCA
 4101  CGAGATTCTT CGCCCTCCGA GAGCTGCATC AGGTCGGAGA CGCTGTCGAA
 4151  CTTTTCGATC AGAAACTTCT CGACAGACGT CGCGGTGAGT TCAGGCTTTT
 4201  TCATCACTAG ACTTCGTTGC AGTTGAGTTT CTGTTTTCGG GCTTGTTGGT
 4251  ATACTTGTTT GATTAATGTG GCTGATAGCT AAGCTATCGC AAATATTTGT
 4301  ACAAGACATG AGGGGGCAAA AAAATAAGCC AAGTGTATTT GTTAGGCTTC
 4351  CGTGCCTGTT AGAGAAATAT GTAAGGAAAC GCGTGCGCTA CACAGCTTGA
 4401  GCACCCATCG AATTGGGACG AGAACATGTT AAACTCAGTA CTCACCCGAT
 4451  AACCCCTATT TTCAATGCAC GCCCACCCAG AGACACGGAA TAGCATAAAA
 4501  CAACCCCATC ACTGCCACAA AATGGACATT TTGCACAAAA CGCACTATTG
 4551  ACTAGTATTC GACTATACTA ATAAGAGATA CCTATCAGAC ACGATATACT
 4601  GTGAAAAGGA CATTAAGCTC ACTATGCATC TGAAAACGCT TTCACCTCAC
 4651  AACCTACTTA CACACACCTT TCCGCTTAAC CCGCTAGCTT TCCACCCAGC
 4701  GCGGGTGCAT TCTGGCTCTT ATATATACTT ATTGTCATGA CAGAGTATAT
 4751  TGTACTGTGT TGATAAGGGA CGGGTAACTG TATTGAAGAG CCGATGCTTT
 4801  TGACATGTTA GATATAATAT GTTTTATTGT AAAGTCAATA CAACACACAA
 4851  TAGGATAATA ATGATAAAGT TAAAAAAGTA TATATAGTAA TAGAAATATA
 4901  TCTTATATAG GAAAGATTAA GCAGTAAAAG TAGCGCTTAC GGCGTACACC
 4951  CTATCAATGA TAGAGTGGTA CCCTGCACGC GCCTTCGAGT TTTTTTTCCT
 5001  TTTCCCCATT TTTTTCAACT TGAAGACTTC AATTACACCA AAAAGTAAAA
 5051  TTCACAAGCT TNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5101  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5151  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5201  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5251  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5301  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5351  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5401  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5451  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5501  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5551  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5601  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5651  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5701  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5751  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5801  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5851  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5901  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 5951  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6001  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6051  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6101  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6151  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6201  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6251  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6301  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6351  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6401  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6451  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6501  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6551  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6601  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6651  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6701  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6751  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6801  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6851  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6901  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 6951  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7001  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7051  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7101  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7151  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7201  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7251  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7301  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7351  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7401  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7451  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7501  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN
 7551  NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNGGAT
 7601  CCTGCCCATT TAGTTGGCTT TTCCCTTGTC TCGTGTCTTT TCCGTGGAAA
 7651  GGTTCCCGGA GTAATCTGAT GGCACAGCAG GGAGGTGCGC CTGCAGGTTG
 7701  GTTAGGAAGG GGGGATGATG TAAAAGAAGA AAATGGGGGG ATATCTTATG
 7751  TTTAAGAGGA TAAATAATGT GAAGGGGCTT TATATGCTTG CTTGGTTGTC
 7801  TCGTTGGTGC ATGGGGAATC TGCATGTTTG CTTTGGAGCA CGCGTGGTAC
 7851  ATCAATGGGA TCATATCTTG CTGCCTCCCG CAGCTCACCG TGCGAGCTGC
 7901  CGGGACCCCG TTTCTCATAG GCGTGTGCAC CTTCCGTCTG AGACCTGTAA
 7951  ATGGTTAAAA AGGAATATAT AATTACCTTT TGAAAAGTGG TAAAAAACGA
 8001  ATATATCTTT TTTTTTGGTT TTAATACGCT CTTTTGTGTG TATGAGAGGA
 8051  ATAAATAAGT GTGTGTGTGT GTGTGTGTTG TTGTTGTTGA AAAAATGAAG
 8101  GACACGAAAG CGCTCATTAA GGGTGGGGAT ACACCAGGCA TAATTATCCG
 8151  AAAATATTTT GATGCACCAA ATAAGTGAAC ATACTGACGA AATCAAAGAG
 8201  TTGGAGGATG GATAGGGAGG CCTCATTGGC AGTGGTAAAT TGATTTGACT
 8251  TGAATATGAT ACCCTCTTAC TGTTTCATTG CCACTACAAG TTGGTTTCCT
 8301  TCCCCTGCAG GCATGCAAGC TAGGCCACAC CTATCAGCCG GTGTGTTGCC
 8351  AAAGACATTC TGTATAATTT TCCAACTACC CTAACCTGAT TCACTCATCG
 8401  TTGAGCATTT GTTCTTCTAC TTTTTGACCG CAAGTGTCGT TATTAGTATT
 8451  CGGTCCTGGG CTGCAGGCTG CTAACAAAGC CCGAAAGGAA GCTGAGTTGG
 8501  CTGCTGCCAC CGCTGAGCAA TAACTAGCAT AACCCCTTGG GGCCTCTAAA
 8551  CGGGTCTTGA GGGGTTTTTT GCTGAAAGGA GAACTATATC CGGATAGGGC
 8601  TGCTAACAAA GCCCGAAAGG AAGCTGAGTT GGCTGCTGCC ACCGCTGAGC
 8651  AATAACTAGC ATAACCCCTT GGGGCCTCTA AACGGGTCTT GAGGGGTTTT
 8701  TTGCTGAAAG GAGAACTATA TCCGGATTGG GCCCCTGCAG