Table 1.

Modification of tandem transcription activator-like effector intermediate vectors

Vector	Backbone	Insert type	Subcloning enzymes	Sequence of short oligos or PCR primers
pFusX1*	pFus_A30A	N/A	N/A	N/A
pFusX2	pFus_A30B	Short oligos	AgeI/AatII	pFusX2-S:
				CCGGTGGTCTCTGGCGGCAAGCAAGCGCTCGAAACGGTGCAGCGGCTGTTGCCGGTGCTGTGCCAGGACCATGGCGAGACGT
				pFusX2-AS: CTCGCCATGGTCCTGGCACAGCACCGGCAACAGCCGCTGCACCGTTTCGAGCGCTTGCTTGCCGCCAGAGACCA
pFusX3	pFus_A30B	PCR	AflII/AatII	pFusX3-F: ATTCTTAAGCGTCTCCATGGCCTGACCCCG
				pFusX3-R: ATTGACGTCTCGCAGGCCATGGTCCTGG
pFusX4	pFus_A30B	PCR	AflII/AatII	pFusX4-F: GTTCTTAAGCGTCTCCCCTGACCCCG
				pFusX4-R: GTTGACGTCTCGGCCATGGTCCTGG

Shown are restriction enzymes and sequences of short oligos or primers used in modification of pFus_A30B into pFusX2, pFusX3, and pFusX4. For pFusX2, annealed reverse complementary short oligos are used directly as the insert. For pFusX3 and pFusX4, PCR products from primers with pFus_A30B as template are used as inserts. No modification is needed for pFusX1, which is identical to pFus_A30B.

N/A, not applicable.