Part:BBa_K277121

3L.3_23.D2.17

3L.3_23.D2.17 is 505 bases long and is cloned into the pGem-T vector.

3L.3_23.D2.17 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. The whole synthetic chromosome may be viewed at http://macbeth.clark.jhu.edu/cgi-bin/gbrowse 3L.3_23.D2.17 is a constituent of 3L.3_23.D2 (along with 3L.3_23.D2.01, 3L.3_23.D2.02, 3L.3_23.D2.03, 3L.3_23.D2.04, 3L.3_23.D2.05, 3L.3_23.D2.06, 3L.3_23.D2.07, 3L.3_23.D2.08, 3L.3_23.D2.09, 3L.3_23.D2.10, 3L.3_23.D2.11, 3L.3_23.D2.12, 3L.3_23.D2.13, 3L.3_23.D2.14, 3L.3_23.D2.15, and 3L.3_23.D2.16.)

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

kind and name offset notes

gene YCL005W-A (72..293) Vacuolar H+ ATPase subunit e of the V0 (membrane) sector%2C essential for vacuolar acidification

loxP_site loxPsym_YCL005W-A (297..330)

gene YCL007C (23..242) Dubious ORF unlikely to encode a protein%3B overlaps verified ORF YCL005W-A%3B mutations in YCL007C were thought to confer sensitivity to calcofluor white%2C but this phenotype was later shown to be due to the defect in YCL005W-A

Sequence (corresponds to coordinates 88015..88519 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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 272