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.

Landmarks
0001-0592: (600) RRNA targeting fragment SacI
0593-3118: (2818) Vector backbone (pUC/pGEM)
3119-3410: (292) T. brucei actin mRNA processing 3′
3411-4022: (612) Hygromycin B phosphotransferase ORF ΔSTART SalI
4051-4365: (315) Expression Site promotor
4377-4615: (239) T. brucei tubulin mRNA processing 5′
4638-5237: (600) Puromycin acetyl transferase ORF
5248-5570: (323) 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 ATATCGACGC TCTCCCTTAT
 2851  GCGACTCCTG CATTAGGAAG CAGCCCAGTA GTAGGTTGAG GCCGTTGAGC
 2901  ACCGCCGCCG CAAGGAATGG TGCAAGGAGA TGGCGCCCAA CAGTCCCCCG
 2951  GCCACGGGGC CTGCCACCAT ACCCACGCCG AAACAAGCGC TCATGAGCCC
 3001  GAAGTGGCGA GCCCGATCTT CCCCATCGGT GATGTCGGCG ATATAGGCGC
 3051  CAGCAACCGC ACCTGTGGCG CCGGTGATGC CGGCCACGAT GCGTCCGGCG
 3101  TAGAGGATCT GGCTAGGAAT ACTGCATAGA TAACAAACGC ATCAAACACA
 3151  ACTGGTGGCA CTATTTCAAT CATGTCGACA CACCAAGACG AGAAAATTTG
 3201  AGTTAAACAA GATGCCTACA ACCCTATATT TATTATCTTC AAGTGATCTT
 3251  CTCTTCTCTT GTGCGCTGTA CGTAAATGTG TTGCAAATAA GGCATAGGGC
 3301  TGAGTACAGG CACCACACGC GAGGAAAATC ATAAGCACTA GCCGTGTCAA
 3351  CTCACAGCGA CTACAGAACA ATTTTGGCCA CACAACCCGG TGTTAGGATC
 3401  TCCGAGGCCT CTATTCCTTT GCCCTCGGAC GAGTGCTGGG GCGTCGGTTT
 3451  CCACTATCGG CGAGTACTTC TACACAGCCA TCGGTCCAGA CGGCCGCGCT
 3501  TCTGCGGGCG ATTTGTGTAC GCCCGACAGT CCCGGCTCCG GATCGGACGA
 3551  TTGCGTCGCA TCGACCCTGC GCCCAAGCTG CATCATCGAA ATTGCCGTCA
 3601  ACCAAGCTCT GATAGAGTTG GTCAAGACCA ATGCGGAGCA TATACGCCCG
 3651  GAGCCGCGGC GATCCTGCAA GCTCCGGATG CCTCCGCTCG AAGTAGCGCG
 3701  TCTGCTGCTC CATACAAGCC AACCACGGCC TCCAGAAGAA GATGTTGGCG
 3751  ACCTCGTATT GGGAATCCCC GAACATCGCC TCGCTCCAGT CAATGACCGC
 3801  TGTTATGCGG CCATTGTCCG TCAGGACATT GTTGGAGCCG AAATCCGCGT
 3851  GCACGAGGTG CCGGACTTCG GGGCAGTCCT CGGCCCAAAG CATCAGCTCA
 3901  TCGAGAGCCT GCGCGACGGA CGCACTGACG GTGTCGTCCA TCACAGTTTG
 3951  CCAGTGATAC ACATGGGGAT CAGCAATCGC GCATATGAAA TCACGCCATG
 4001  TAGTGTATTG ACCGATTCCT TGCGGTCGGC CGCTCTAGAA CTAGTGGATC
 4051  AACTGTGCTC TCGGAGAGAG AGAGGGTGGG ATTTTTTTCT TTAAATAAAG
 4101  GGAGATGTAT TCGGAGGAGT TGCATTAACG TTGTCCCATG TTTTACAAAA 
 4151  AACGAGTCTG AAAGAACGCC GACTCAATGG CCATCCAAGC GGAATAACCT 
 4201  TCTAAAAGAA TCATATCCCT ATTACCACAC CAGTTTATAT TACAGGGGAG
 4251  GTTATTACAG AAATCTAGGA TATCAGACTC AAGGTGTGGC GGACGTCTCG
 4301  AACCGATCGT GAGTTGGCCT GAGCGTGCCG TCGTTACCGG AGTGCCCGTA
 4351  TTCGATTTAT GTCGAGATCC TCTAGATGGG TCCCATTGTT TGCCTCTTCC
 4401  GCTGTGTGGA GTGCGCCTAC ACGCACTTCT CACTTCGTAA GTGGTGGTGG
 4451  CGTAAGTATT GCCTAATGTT GACTCTATGT TCTCCTCTCC TCACCCCCTC
 4501  GCGGTGCTGA TTTCTGACAG ATCTTCAAAC ACTAGTTTAA GCAAAGGACT
 4551  ATTCATCCGT TTATATTAGC AACAGTAGGT ACTAGCACCA CTAACAACAA
 4601  CAACAAAGCA CTTCTGCAGG AATTCGATAT CAAGCTTATG ACCGAGTACA
 4651  AGCCCACGGT GCGCCTCGCC ACCCGCGACG ACGTCCCCCG GGCCGTACGC
 4701  ACCCTCGCCG CCGCGTTCGC CGACTACCCC GCCACGCGCC ACACCGTCGA
 4751  CCCGGACCGC CACATCGAGC GGGTCACCGA GCTGCAAGAA CTCTTCCTCA
 4801  CGCGCGTCGG GCTCGACATC GGCAAGGTGT GGGTCGCGGA CGACGGCGCC
 4851  GCGGTGGCGG TCTGGACCAC GCCGGAGAGC GTCGAAGCGG GGGCGGTGTT
 4901  CGCCGAGATC GGCCCGCGCA TGGCCGAGTT GAGCGGTTCC CGGCTGGCCG
 4951  CGCAGCAACA GATGGAAGGC CTCCTGGCGC CGCACCGGCC CAAGGAGCCC
 5001  GCGTGGTTCC TGGCCACCGT CGGCGTCTCG CCCGACCACC AGGGCAAGGG
 5051  TCTGGGCAGC GCCGTCGTGC TCCCCGGAGT GGAGGCGGCC GAGCGCGCCG
 5101  GGGTGCCCGC CTTCCTGGAG ACCTCCGCGC CCCGCAACCT CCCCTTCTAC
 5151  GAGCGGCTCG GCTTCACCGT CACCGCCGAC GTCGAGGTGC CCGAAGGACC
 5201  GCGCACCTGG TGCATGACCC GCAAGCCCGG TGCCTGAGAA TTCGCCCCTA
 5251  GAAAGTGTGA CAACGTCGCA CCATGTGTAG GTTTTCATTT ATGTTCTTTC
 5301  TTTCTTTCTT TTGTGAATTT GTTTTCTGTC TCAAATGTTT TTAATTCGCT
 5351  TGGGACCTAT GTTTTTCTTG TTTTTTTGCT CACCCTTTGT GTAGGAGGCA
 5401  CCCTGTCACG TCTGTGGTTG CGTGTATGCC TTCCTTCCCC TTATTCGCTT
 5451  CTTCCTGTCG TGTCACACCT CTTTCTCCCT CTCCCTTTCC GCCTTTTCTT
 5501  TCAATCTTGT TTTCTCGACC AGCCCTACTA GAGGAGAAAG AATAGTAACC
 5551  CTTTCATCAA AGAAAATAGT CGAGGGGGGG CCCCTGCAG