IGEM:MIT/2005/Primers

The melting temperatures were calculated mostly by the following site: Oligonucleotide Primer Check


 * scFv (pCT-4-4-20, pCT302 S101A, pCT-4M5.3 anti-fluorescein plasmids)

FWD: 5' CATCAT GAATTC GCGGCCGC T TCTAGA GACGTCGTTATGACTCAAACACC 3' T_m,i=71.8 T_m,f=88.5 rand  EcoRI   NotI      XbaI       Primer

REV: 5' GCTCAT ACTAGT TTATTA CTTATCATCATCATCCTTATAATC GGAGACGGTGACTGAGGTTCC 3' T_m,i=74.7 T_m,f=87.1 rand   SpeI   Stop          FLAG Tag                 Primer

fecA-Fwd 5’CATCAT  GAATTC  GCGGCCGC  TCTAGA TGACGCCGTTACGCGTTTTTCG 3’ T_m,i=73.2 T_m,f=89.6 rand   EcoRI    NotI    XbaI     primer fecA-Rev 5’GCCATA  ACTAGT  TTATTA  TCAGAACTTCAACGACCCCT 3’ T_m,i=68.2 T_m,f=79.9 rand   SpeI     stop    primer
 * FecA

Middle Fwd- 5'GGACGGCAGCGGACTGCAAGTAAAACCGCTGGGAAATAAC 3' T_m=84.7 Middle Rev- 5’GTTATTTCCCAGCGGTTTTACTTGCAGTCCGCTGCCGTCC 3' T_m=84.7

fecA Promoter Fwd- 5’CATCAT GAATTC  GCGGCCGC T TCTAGA  TGGATAAACATTTCACCACTG 3' T_m,i=65.0 T_m,f=86.2 rand   EcoRI    NotI      XbaI     primer fecA Promoter Rev- 5'GCCATA CTGCAG  CGGCCGC  T ACTAGT  CACAAGTCGATACTCAGCTTG 3' T_m,i=68.9 T_m,f=89.8 rand   PstI     NotI      SpeI     primer fecR-Fwd 5' CATCAT GAATTC  GCGGCCGC TCTAGA tgaatcctttgttaaccgattccc 3'T_m 66 rand  EcoRI     NotI     XbaI   primer fecR-Rev 5' GCCATA ACTAGT  TTATTA ttacagtggtgaaatgtttatccag 3' T_m 68 rand  SpeI     stop   primer Middle Fwd 5'gtgaaagcctacagttcagcgcctcag3' T_m 84 Middle Rev 5'ctgaggcgctgaactgtaggctttcac3' T_m 84
 * FecR

fecI-Fwd 5'CATCAT GAATTC GCGGCCGC T TCTAGA tgtctgaccgcgccactacc3' T_m 68 rand  EcoRI    NotI     XbaI  primer fecI-Rev 5'GCCATA CTGCAG CGGCCGC T ACTAGT A TTATTA tcataacccatactccagacggaa3' T_m 70 rand PstI    NotI      SpeI     stop  primer
 * FecI
 * MalE (updated/fixed)

fwd: 5' CCGTTA GAATTC GCGGCCGC T TCTAGA atgaaaataaaaacaggtgcacgc3' T_m,i=68.8 T_m,f=85.5 junk   EcoRI    NotI     XbaI       Primer * rev: 5' ACTGAG CTGCAG GCGGCCGC T ACTAGT TTATTA  TTACTTGGTGATACGAGTCTGC 3' T_m,i=69.5 T_m,f=89.4 junk   PstI    NotI      SpeI   Stop        Primer

fwd: 5'    CATGAG  GAATTC GCGGCCGC T TCTAGA ATGAAACAGAGCACCATTGCG 3' T_m,i=68.9 T_m,f=88.8 junk   EcoRI    NotI     XbaI       Primer rev: 5'    AAGCTA  CTGCAG GCGGCCGC T ACTAGT TTATTA TTATTTCAGGCCCAGCGCC 3' T_m,i=69.7 T_m,f=89.8 junk   PstI    NotI      SpeI   Stop        Primer
 * PhoA

Middle(1) fwd: 5' caccgcggaactgcaagatgcgaccccggcg 3' T_m=88.8 * rev: 5' CGCCGGGGTCGCATCTTGCAGTTCCGCGGTG 3' T_m=88.8 *

Middle(2) fwd: 5' aggcttttttctgcaagtggaaggcgcgagc 3' T_m=82.2 * rev: 5' GCTCGCGCCTTCCACTTGCAGAAAAAAGCCT 3' T_m=82.2 *