Stem loop RNAi cassettes

Cassette sequenced in full, February 2008

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

All of the restriction sites listed are unique.

MCS1/2

KpnI 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 ApaI
MCS3/4
KpnI 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 ApaI