Part:BBa_K277057

3L.3_23.B3.05

3L.3_23.B3.05 is 774 bases long and is cloned into the pGem-T vector.

3L.3_23.B3.05 was designed as a piece of synthetic chromosome 3 with the goal of minimizing and stabilizing that chromosome and to that end has had any tRNAs, introns, repeat regions, and transposons that were present in the wildtype chromosome removed. In addition a very few gene sequences were slightly recoded to add or remove restriction enzyme recognition sites to facilitate assembly; most gene sequences were slightly recoded to introduce unique primers for diagnostic PCR amplification, and some gene sequences were slightly recoded to address the distribution of stop codon usage. 3L.3_23.B3.05 is a constituent of 3L.3_23.B3 (along with 3L.3_23.B3.01, 3L.3_23.B3.02, 3L.3_23.B3.03, 3L.3_23.B3.04, 3L.3_23.B3.06, 3L.3_23.B3.07, 3L.3_23.B3.08, and 3L.3_23.B3.09.)

This part contains at least part of the following features (positions offset from first base of sequence):

kind and name offset notes

mutation_affecting_coding_sequence YCL040W_re_remove_MmeI (127..138) removal of MmeI

reverse_primer YCL040W_tagr1v1 (699..723)

mutation_affecting_coding_sequence YCL040W_re_remove_Bpu10I (79..87) removal of Bpu10I

gene YCL040W (-530..+972) Glucokinase%2C catalyzes the phosphorylation of glucose at C6 in the first irreversible step of glucose metabolism%3B one of three glucose phosphorylating enzymes%3B expression regulated by non-fermentable carbon sources

mutation_affecting_coding_sequence YCL040W_re_remove_BpuEI (358..366) removal of BpuEI

forward_primer YCL040W_tagf1v1 (405..432)

forward_primer YCL040W_tagf2v1 (567..594)

Sequence (the first 774 bases correspond to coordinates 41708..42481 in synthetic chromosome yeast_chr3_3_23)

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 223
  
• 1000
  COMPATIBLE WITH RFC[1000]