The Primer Generator

Original nucleotide sequence	AAATTTGGCACA
Original amino acid sequence	KFGT
Desired amino acid sequence	KYGT
Use of restriction enzymes with site size less than 5 bp	No
Use of only most common restriction enzymes	No
Maximum number of substitutions compared to original sequence	3

The following primers have been generated:

Click on enzyme name to access corresponding entry in The Restriction Enzyme Database (REBASE)
Positions of cut sites are calculated relative to the beginning of the primer.
Differences between primer sequence and original nucleotide sequence are bolded

Primer Sequence	Deleted Restriction Sites	Cut Positions	New Restriction Sites	Cut Positions	Number of Substitutions
AAATATGGCACA	AcsI	1, 5	None		1
AAATATGGCACA	ApoI	1, 5			1
AAATATGGCACC	AcsI	1, 5	AccB1I	7, 11	2
	ApoI	1, 5	BanI	7, 11
			BshNI	7, 11
			Eco64I	7, 11
AAATATGGGACA	AcsI	1, 5	BsmFI	23	2
AAATATGGGACA	ApoI	1, 5			2
AAATATGGAACA	AcsI	1, 5	None		2
AAATATGGAACA	ApoI	1, 5			2
AAGTATGGCACA	AcsI	1, 5	None		2
AAGTATGGCACA	ApoI	1, 5			2
AAATATGGCACG	AcsI	1, 5	None		2
AAATATGGCACG	ApoI	1, 5			2
AAATACGGCACA	AcsI	1, 5	None		2
AAATACGGCACA	ApoI	1, 5			2
AAATATGGTACA	AcsI	1, 5	None		2
AAATATGGTACA	ApoI	1, 5			2
AAATATGGCACT	AcsI	1, 5	None		2
AAATATGGCACT	ApoI	1, 5			2
AAATATGGTACC	AcsI	1, 5	Acc65I	7, 11	3
	ApoI	1, 5	AccB1I	7, 11
			Asp718I	7, 11
			BanI	7, 11
			BshNI	7, 11
			Eco64I	7, 11
			KpnI	11, 7
AAATATGGGACC	AcsI	1, 5	AvaII	8, 11	3
	ApoI	1, 5	Bme18I	8, 11
			BsmFI	23
			Eco47I	8, 11
			HgiEI	8, 11
			SinI	8, 11
AAGTATGGCACC	AcsI	1, 5	AccB1I	7, 11	3
	ApoI	1, 5	BanI	7, 11
			BshNI	7, 11
			Eco64I	7, 11
AAATACGGCACC	AcsI	1, 5	AccB1I	7, 11	3
	ApoI	1, 5	BanI	7, 11
			BshNI	7, 11
			Eco64I	7, 11
AAGTATGGGACA	AcsI	1, 5	BsmFI	23	3
AAGTATGGGACA	ApoI	1, 5			3
AAATATGGGACG	AcsI	1, 5	BsmFI	23	3
AAATATGGGACG	ApoI	1, 5			3
AAATACGGGACA	AcsI	1, 5	BsmFI	23	3
AAATACGGGACA	ApoI	1, 5			3
AAATATGGGACT	AcsI	1, 5	BsmFI	23	3
AAATATGGGACT	ApoI	1, 5			3
AAATATGGAACC	AcsI	1, 5	None		3
AAATATGGAACC	ApoI	1, 5			3
AAATACGGTACA	AcsI	1, 5	None		3
AAATACGGTACA	ApoI	1, 5			3
AAATACGGCACT	AcsI	1, 5	None		3
AAATACGGCACT	ApoI	1, 5			3
AAGTATGGAACA	AcsI	1, 5	None		3
AAGTATGGAACA	ApoI	1, 5			3
AAATATGGTACT	AcsI	1, 5	None		3
AAATATGGTACT	ApoI	1, 5			3
AAATATGGAACG	AcsI	1, 5	None		3
AAATATGGAACG	ApoI	1, 5			3
AAGTATGGCACG	AcsI	1, 5	None		3
AAGTATGGCACG	ApoI	1, 5			3
AAATACGGAACA	AcsI	1, 5	None		3
AAATACGGAACA	ApoI	1, 5			3
AAATATGGAACT	AcsI	1, 5	None		3
AAATATGGAACT	ApoI	1, 5			3
AAGTACGGCACA	AcsI	1, 5	None		3
AAGTACGGCACA	ApoI	1, 5			3
AAATACGGCACG	AcsI	1, 5	None		3
AAATACGGCACG	ApoI	1, 5			3
AAGTATGGTACA	AcsI	1, 5	None		3
AAGTATGGTACA	ApoI	1, 5			3
AAATATGGTACG	AcsI	1, 5	None		3
AAATATGGTACG	ApoI	1, 5			3
AAGTATGGCACT	AcsI	1, 5	None		3
AAGTATGGCACT	ApoI	1, 5			3

Authors:

	Joseph Lawler
	Alexander Turchin