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