Parts Created
From 2006.igem.org
Hin enhancer with biobrick ends:
gaattcgcggccgcttctagattcgggtgtcaacaattgaccaaaatatcgatttacagcgtaatgcgctttctagtgcaaattgtgaccgcattttactagttgcggccgcctgcag
- Changed the first base in bold from and "A" to a "T" to avoid the creation of a SpeI site within the enhancer which would lead to a problem when cutting the DNA because the biobricks contain a SpeI site.
Hin PCR Primers:
Left primer including biobrickends (5'->3'):
gcattctagaatggctactattgggtatattcgggtgtcaacaattgaccaaaatatcgatttacagcgtaatgcgcttacCagtgcaaattg
- The base in bold has been mutated from a "T" to a "C" to eliminate a SpeI site. Because the Hin gene codes for a protein the base substitution was chosen so that the amino acid that will form will be the same as that formed from the wild type gene.
Right Primer including biobick ends (5'-> 3'):
atgcctgcaggcggccgcactagtttaattcattcgtttttttatac
Oligoes used to make the recombinational enhancer
AATTCGCGGCCGCTTCTAGATTCGGGTGTCAACAATTGACCAAAATAT
CGATTTACAGCGTAATGCGCTTTCTAGTGCAAATTGTGACCGC
ATTTTACTAGTTGCGGCCGCCTGCA
GGCGGCCGCAACTAGTAAAATGCGGTCACAATTTGCACTAGAA
AGCGCATTACGCTGTAAATCGATATTTTGGTCAATTGTTGACACCCGAATCTAGAAGCGGCCGCG