ph3Ep
Designed to replace a pRP expression construct previously integrated at a RRNA spacer locus, generating a ‘landing pad‘ for future pRPa integrations.
Linearise with SacI/SalI prior to transfection.
Derived from known sequence and junction sequencing, July 2005.
NB: Follow-up sequencing in October 2017 identified two single base errors in the PAC ORF and missing flanking sequence (3-bp upstream and 59-bp downstream of PAC). The sequence below has been corrected, accordingly.
| Landmarks | |
| 0007-0580: (574) | RRNA targeting fragment SacI |
| 0593-3118: (2818) | Vector backbone (pUC/pGEM) |
| 3119-3410: (292) | T. brucei actin mRNA processing 3′ SalI |
| 3410-4025: (616) | Hygromycin B phosphotransferase ORF ΔSTART |
| 4053-4359: (307) | Expression Site promotor |
| 4376-4612: (237) | T. brucei tubulin mRNA processing 5′ |
| 4640-5239: (600) | Puromycin acetyl transferase ORF |
| 5305-5637: (333) | T. brucei tubulin mRNA processing 3′ |
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 GAGCTCGAAT TCTGCATTAA 601 TGAATCGGCC AACGCGCGGG GAGAGGCGGT TTGCGTATTG GGCGCTCTTC 651 CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG 701 CGGTATCAGC TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG 751 GATAACGCAG GAAAGAACAT GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA 801 CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT CCATAGGCTC CGCCCCCCTG 851 ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG AAACCCGACA 901 GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC 951 TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT 1001 CGGGAAGCGT GGCGCTTTCT CATAGCTCAC GCTGTAGGTA TCTCAGTTCG 1051 GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT GTGCACGAAC CCCCCGTTCA 1101 GCCCGACCGC TGCGCCTTAT CCGGTAACTA TCGTCTTGAG TCCAACCCGG 1151 TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA CAGGATTAGC 1201 AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA 1251 CTACGGCTAC ACTAGAAGGA CAGTATTTGG TATCTGCGCT CTGCTGAAGC 1301 CAGTTACCTT CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC 1351 ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC AAGCAGCAGA TTACGCGCAG 1401 AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT CTTTTCTACG GGGTCTGACG 1451 CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAT GAGATTATCA 1501 AAAAGGATCT TCACCTAGAT CCTTTTAAAT TAAAAATGAA GTTTTAAATC 1551 AATCTAAAGT ATATATGAGT AAACTTGGTC TGACAGTTAC CAATGCTTAA 1601 TCAGTGAGGC ACCTATCTCA GCGATCTGTC TATTTCGTTC ATCCATAGTT 1651 GCCTGACTCC CCGTCGTGTA GATAACTACG ATACGGGAGG GCTTACCATC 1701 TGGCCCCAGT GCTGCAATGA TACCGCGAGA CCCACGCTCA CCGGCTCCAG 1751 ATTTATCAGC AATAAACCAG CCAGCCGGAA GGGCCGAGCG CAGAAGTGGT 1801 CCTGCAACTT TATCCGCCTC CATCCAGTCT ATTAATTGTT GCCGGGAAGC 1851 TAGAGTAAGT AGTTCGCCAG TTAATAGTTT GCGCAACGTT GTTGCCATTG 1901 CTACAGGCAT CGTGGTGTCA CGCTCGTCGT TTGGTATGGC TTCATTCAGC 1951 TCCGGTTCCC AACGATCAAG GCGAGTTACA TGATCCCCCA TGTTGTGCAA 2001 AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT CGTTGTCAGA AGTAAGTTGG 2051 CCGCAGTGTT ATCACTCATG GTTATGGCAG CACTGCATAA TTCTCTTACT 2101 GTCATGCCAT CCGTAAGATG CTTTTCTGTG ACTGGTGAGT ACTCAACCAA 2151 GTCATTCTGA GAATAGTGTA TGCGGCGACC GAGTTGCTCT TGCCCGGCGT 2201 CAATACGGGA TAATACCGCG CCACATAGCA GAACTTTAAA AGTGCTCATC 2251 ATTGGAAAAC GTTCTTCGGG GCGAAAACTC TCAAGGATCT TACCGCTGTT 2301 GAGATCCAGT TCGATGTAAC CCACTCGTGC ACCCAACTGA TCTTCAGCAT 2351 CTTTTACTTT CACCAGCGTT TCTGGGTGAG CAAAAACAGG AAGGCAAAAT 2401 GCCGCAAAAA AGGGAATAAG GGCGACACGG AAATGTTGAA TACTCATACT 2451 CTTCCTTTTT CAATATTATT GAAGCATTTA TCAGGGTTAT TGTCTCATGA 2501 GCGGATACAT ATTTGAATGT ATTTAGAAAA ATAAACAAAT AGGGGTTCCG 2551 CGCACATTTC CCCGAAAAGT GCCACCTGAC GTCTAAGAAA CCATTATTAT 2601 CATGACATTA ACCTATAAAA ATAGGCGTAT CACGAGGCCC TTTCGTCTCG 2651 CGCGTTTCGG TGATGACGGT GAAAACCTCT GACACATGCA GCTCCCGGAG 2701 ACGGTCACAG CTTGTCTGTA AGCGGATGCC GGGAGCAGAC AAGCCCGTCA 2751 GGGCGCGTCA GCGGGTGTTG GCGGGTGTCG GGGCTGGCTT AACTATGCGG 2801 CATCAGAGCA GATTGTACTG AGAGTGCACC ATTCGACGCT CTCCCTTATG 2851 CGACTCCTGC ATTAGGAAGC AGCCCAGTAG TAGGTTGAGG CCGTTGAGCA 2901 CCGCCGCCGC AAGGAATGGT GCAAGGAGAT GGCGCCCAAC AGTCCCCCGG 2951 CCACGGGGCC TGCCACCATA CCCACGCCGA AACAAGCGCT CATGAGCCCG 3001 AAGTGGCGAG CCCGATCTTC CCCATCGGTG ATGTCGGCGA TATAGGCGCC 3051 AGCAACCGCA CCTGTGGCGC CGGTGATGCC GGCCACGATG CGTCCGGCGT 3101 AGAGGATCTG GCTAGGAATA CTGCATAGAT AACAAACGCA TCAAACACAA 3151 CTGGTGGCAC TATTTCAATC ATGTCGACAC ACCAAGACGA GAAAATTTGA 3201 GTTAAACAAG ATGCCTACAA CCCTATATTT ATTATCTTCA AGTGATCTTC 3251 TCTTCTCTTG TGCGCTGTAC GTAAATGTGT TGCAAATAAG GCATAGGGCT 3301 GAGTACAGGC ACCACACGCG AGGAAAATCA TAAGCACTAG CCGTGTCAAC 3351 TCACAGCGAC TACAGAACAA TTTTGGCCAC ACAACCCGGT GTTAGGATCT 3401 CCGAGGCCTC TATTCCTTTG CCCTCGGACG AGTGCTGGGG CGTCGGTTTC 3451 CACTATCGGC GAGTACTTCT ACACAGCCAT CGGTCCAGAC GGCCGCGCTT 3501 CTGCGGGCGA TTTGTGTACG CCCGACAGTC CCGGCTCCGG ATCGGACGAT 3551 TGCGTCGCAT CGACCCTGCG CCCAAGCTGC ATCATCGAAA TTGCCGTCAA 3601 CCAAGCTCTG ATAGAGTTGG TCAAGACCAA TGCGGAGCAT ATACGCCCGG 3651 AGCCGCGGCG ATCCTGCAAG CTCCGGATGC CTCCGCTCGA AGTAGCGCGT 3701 CTGCTGCTCC ATACAAGCCA ACCACGGCCT CCAGAAGAAG ATGTTGGCGA 3751 CCTCGTATTG GGAATCCCCG AACATCGCCT CGCTCCAGTC AATGACCGCT 3801 GTTATGCGGC CATTGTCCGT CAGGACATTG TTGGAGCCGA AATCCGCGTG 3851 CACGAGGTGC CGGACTTCGG GGCAGTCCTC GGCCCAAAGC ATCAGCTCAT 3901 CGAGAGCCTG CGCGACGGAC GCACTGACGG TGTCGTCCAT CACAGTTTGC 3951 CAGTGATACA CATGGGGATC AGCAATCGCG CATATGAAAT CACGCCATGT 4001 AGTGTATTGA CCGATTCCTT GCGGTCGGCC GCTCTAGAAC TAGTGGATCA 4051 ACTGTGCTCT CGGAGAGAGA GAGGGTGGGA TTTTTTTCTT TAAATAAAGG 4101 GAGATGTATT CGGAGGAGTT GCATTAACGT TGTCCCATGT TTTACAAAAA 4151 ACGAGTCTGA AAGAACGCCG ACTCAATGGC CATCCAAGCG GAATAACCTT 4201 CTAAAAGAAT CATATCCCTA TTACCACACC AGTTTATATT ACAGGGGAGG 4251 TTATTACAGA AATCTAGGAT ATCAGACTCA AGGTGTGGCG GACGTCTCGA 4301 ACCGATCGTG AGTTGGCCTG AGCGTGCCGT CGTTACCGGA GTGCCCGTAT 4351 TCGATTTATG TCGAGATCCT CTAGATGGGT CCCATTGTTT GCCTCTTCCG 4401 CTGTGTGGAG TGCGCCTACA CGCACTTCTC ACTTCGTAAG TGGTGGTGGC 4451 GTAAGTATTG CCTAATGTTG ACTCTATGTT CTCCTCTCCT CACCCCCTCG 4501 CGGTGCTGAT TTCTGACAGA TCTTCAAACA CTAGTTTAAG CAAAGGACTA 4551 TTCATCCGTT TATATTAGCA ACAGTAGGTA CTAGCACCAC TAACAACAAC 4601 AACAAAGCAC TTCTGCAGGA ATTCGATATC AAGCTTACCA TGACCGAGTA 4651 CAAGCCCACG GTGCGCCTCG CCACCCGCGA CGACGTCCCC AGGGCCGTAC 4701 GCACCCTCGC CGCCGCGTTC GCCGACTACC CCGCCACGCG CCACACCGTC 4751 GATCCGGACC GCCACATCGA GCGGGTCACC GAGCTGCAAG AACTCTTCCT 4801 CACGCGCGTC GGGCTCGACA TCGGCAAGGT GTGGGTCGCG GACGACGGCG 4851 CCGCGGTGGC GGTCTGGACC ACGCCGGAGA GCGTCGAAGC GGGGGCGGTG 4901 TTCGCCGAGA TCGGCCCGCG CATGGCCGAG TTGAGCGGTT CCCGGCTGGC 4951 CGCGCAGCAA CAGATGGAAG GCCTCCTGGC GCCGCACCGG CCCAAGGAGC 5001 CCGCGTGGTT CCTGGCCACC GTCGGCGTCT CGCCCGACCA CCAGGGCAAG 5051 GGTCTGGGCA GCGCCGTCGT GCTCCCCGGA GTGGAGGCGG CCGAGCGCGC 5101 CGGGGTGCCC GCCTTCCTGG AGACCTCCGC GCCCCGCAAC CTCCCCTTCT 5151 ACGAGCGGCT CGGCTTCACC GTCACCGCCG ACGTCGAGGT GCCCGAAGGA 5201 CCGCGCACCT GGTGCATGAC CCGCAAGCCC GGTGCCTGAC GCCCGCCCCA 5251 CGACCCGCAG CGCCCGACCG AAAGGAGCGC ACGACCCCAT GCATCGATGA 5301 ATTCGCCCTA GAAAGTGTGA CAACGTCGCA CCATGTGTAG GTTTTCATTT 5351 ATGTTCTTTC TTTCTTTCTT TTGTGAATTT GTTTTCTGTC TCAAATGTTT 5401 TTAATTCGCT TGGGACCTAT GTTTTTCTTG TTTTTTTGCT CACCCTTTGT 5451 GTAGGAGGCA CCCTGTCACG TCTGTGGTTG CGTGTATGCC TTCCTTCCCC 5501 TTATTCGCTT CTTCCTGTCG TGTCACACCT CTTTCTCCCT CTCCCTTTCC 5551 GCCTTTTCTT TCAATCTTGT TTTCTCGACC AGCCCTACTA GAGGAGAAAG 5601 AATAGTAACC CTTTCATCAA AGAAAATAGT CGAGGGGGGG CCCCTGCAG