IGEM:MIT/2008/Notebook/Yogurt/sequences

Primers

Include sequence, usage, temperatures, concentration, start date.

P1: Forward Primer for Promoter

• • foward sequence: CCG CTT CTA GAG TAA TAC GAC TCA CTA TAG GGA ATA CAA GCT ACT TGT TCT TTT TGC ATA CTA CAG ATT AAA GAG GAG AAA TAC TAG ATG TAC GTC TTC TTT TTT AGG CG
  • length = 110 bp
  • melting temp = 68.8 C
  • GC content % = 37.3

P2: Reverse Primer for Peptide

• • reverse sequence: GTT CTT CTC CTT TAC GCA TAG AAC CAC CAC CAG AAC CAC C
  • length = 69 bp
  • melting temp = 71.8 C
  • GC content % = 50.7

TEV Protease cut site

• Amino Acid Sequence : Glu-Asn-Leu-Tyr-Phe-Gln-Gly (Cuts between Gln-Gly)
• Base Pair Sequence: GAAAACCTGTACTTCCAGGGT

P3: Forward Primer for GFP

forward sequence: GGT GGT TCT GGT GGT GGT TCT ATG CGT AAA GGA GAA GAA C

• • includes 21 bp of 2nd half of linker, and 19 bp of GFP
  • length = 40 bp
  • melting temp = 66.2 C
  • GC content % = 50%

P4: Reverse Primer for GFP

• • reverse sequence: CTG CAG CGG CCG CTA CTA GTA TTA TTA GTG GTG ATG GTG ATG ATG TTT GTA TAG TTC ATC CAT GC
  • length = 65 bp
  • melting temp = 68.5 C
  • GC content % = 44.6

Plasmids

Include sequence, BioBrick #, functional features, digestion map.

PCR Products

Include sequence, template and primers, functional features.

Other

DNA sequence to synthesize - epitope/p1025/linker (6/17/08)

• The signal peptide sequence can be found at the NCBI genome database

• 6/17, updated signal peptide, silent mutations in 1st part of linker (GGC instead of GGT coding for Gly)