SIL082506-Construction of pSB1AK3-J23075

[RA2 Promoter][RBS][trbCf][TT]
Digest PCR product of pSB1A2-J23038 with G00101 and ca641R (BSAI/SpeI, Neb2, ~1600 )
Paste into pSB1AK3-J23028 (XbaI/BsaI, Neb2, Large 5229)
Product is pSB1AK3-J23075.

JCAnderson 20:11, 16 September 2006 (EDT)Revised construction:

Construction of pSB1AK3-J23075

[Pcon][stRBS][trbC][TT]
Digest pSB1AK3-J23028 . . . . . . . . (XbaI/BglI, large-3037 bp)
Digest pBca1020-J23038 . . . . . . . .(SpeI/BglI, small-993bp)
Product is pSB1AK3-J23075

Construction of pSB1AKG0-J23075

[genR]-X-[Pcon][stRBS][trbC][TT]
Digest pSB1AK3-J23028 . . . . . . . . (EcoRI/XbaI, large-3951 bp)
Digest pSB1AG0-J23038 . . . . . . . . (EcoRI/SpeI, small-1538 bp)
Ligate, plate on Amp+Kan+Gen
product is pSB1AKG0-J23075