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