Stem-loop RNAi cassette

Cassettes sequenced in full, February 2008

Two versions are available in the pRPa backbone, containing different multiple cloning sites separated by a 468 bp stuffer (pBluescript 117-585, including 125 bp of the β-galactosidase α-fragment)

NB: The pRP version of the stem-loop RNAi construct is only available with a HYG selectable marker and the MCS1/2 RNAi expression cassette

All of the restriction sites listed are unique.

MCS^1/2

Acc65I SmaI AflII BamHI

   1  GGTACCCGGG CTTAAGGATC CGAGATAGGG TTGAGTGTTG TTCCAGTTTG
  51  GAACAAGAGT CCACTATTAA AGAACGTGGA CTCCAACGTC AAAGGGCGAA
 101  AAACCGTCTA TCAGGGCGAT GGCCCACTAC GTGAACCATC ACCCTAATCA
 151  AGTTTTTTGG GGTCGAGGTG CCGTAAAGCA CTAAATCGGA ACCCTAAAGG
 201  GAGCCCCCGA TTTAGAGCTT GACGGGGAAA GCCGGCGAAC GTGGCGAGAA
 251  AGGAAGGGAA GAAAGCGAAA GGAGCGGGCG CTAGGGCGCT GGCAAGTGTA
 301  GCGGTCACGC TGCGCGTAAC CACCACACCC GCCGCGCTTA ATGCGCCGCT
 351  ACAGGGCGCG TCCATTCGCC ATTCAGGCTG CGCAACTGTT GGGAAGGGCG
 401  ATCGGTGCGG GCCTCTTCGC TATTACGCCA GCTGGCGAAA GGGGGATGTG
 451  CTGCAAGGCG ATTAAGTTGG GTAACGCCAG GGTTTTCTAG AAGCTTCTCG
 501  AGGGCCC

XbaI HindIII XhoI Bsp120I

MCS^3/4

Acc65I AgeI ClaI MfeI SmaI
Note: AgeI & XmaI (isoschizomer of SmaI) give compatible overhangs; ClaI site blocked by Dam methylation

   1  GGTACCGGTA TCGATCAATT GCCCGGGTCC GAGATAGGGT TGAGTGTTGT
  51  TCCAGTTTGG AACAAGAGTC CACTATTAAA GAACGTGGAC TCCAACGTCA
 101  AAGGGCGAAA AACCGTCTAT CAGGGCGATG GCCCACTACG TGAACCATCA
 151  CCCTAATCAA GTTTTTTGGG GTCGAGGTGC CGTAAAGCAC TAAATCGGAA
 201  CCCTAAAGGG AGCCCCCGAT TTAGAGCTTG ACGGGGAAAG CCGGCGAACG
 251  TGGCGAGAAA GGAAGGGAAG AAAGCGAAAG GAGCGGGCGC TAGGGCGCTG
 301  GCAAGTGTAG CGGTCACGCT GCGCGTAACC ACCACACCCG CCGCGCTTAA
 351  TGCGCCGCTA CAGGGCGCGT CCATTCGCCA TTCAGGCTGC GCAACTGTTG
 401  GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT ATTACGCCAG CTGGCGAAAG
 451  GGGGATGTGC TGCAAGGCGA TTAAGTTGGG TAACGCCAGG GTTTTCAGAT
 501  CTTAAGGCCG GCCTCGAGTC TAGAGGGCCC

BglII AflII FseI XhoI XbaI Bsp120I