ThpB

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PCR ThpBF/ ThpBIR on ThpB (610 bp, gp=A)
PCR ThpBR/ ThpBIF on ThpB (712 bp, gp=B)



PCR ThpBAF/ThpBAR on A+B (1285bp BglII/XhoI)
Digest BglII/XhoI
Product pBca9145-BBa I716331



ThpBF Forward BglII site anneals to ThpB ctagaAGATCTgtgaccatcaccccgcccgc
ThpBIF Removing XhoI site in ThpB ggttcgagcggctcctTgaggaccagggctccggac
ThpBIR Removing XhoI site in ThpD gtccggagccctggtcctcAaggagccgctcgaacc
ThpBR Reverse XhoI site anneals to ThpB cgtcactcgagtcaggccgtttcgcggacgc