Team
|
Ethanol (E) or Acetate (A)
|
Gene targeted by CRISPRi gRNA
|
gRNA sequence (without tag at 3' end)
|
Locus targeted (eg. beginning of gene, putative promoter, -35 region)
|
Target template or nontemplate strand
|
red
|
Acetate
|
adhE
|
TTAACGCACTCGTAGAGCGTGTAAA
|
beginning of coding region
|
|
orange
|
Ethanol
|
pta-ack
|
CTATGGCTCCCTGACGTTTT
|
promoter region
|
template strand
|
yellow
|
Ethanol
|
frdA
|
GCTGTGGGATAAAAACAATCTGGAG
|
minus 35 region
|
template strand
|
green
|
E
|
LdhA
|
ttgtgctataaacggcgagtttcat
|
Non template strand
|
In the middle of the gene
|
blue
|
E
|
pta
|
ttcacgacaacgttcaataatcat
|
coding region
|
non-template strand
|
pink
|
Acetate
|
adhE
|
TTACTAAAAAAGTTTAACATTATCA
|
promoter region
|
template strand
|
purple
|
Acetate
|
adhE
|
TTTACTAAAAAAGTTTAACATTATC
|
' -35 region'
|
'template'
|
white
|
Acetate
|
adhE
|
CTGATAATGTTAAACTTTTT
|
promoter
|
non-template strand
|
Team
|
Ethanol (E) or Acetate (A)
|
Gene targeted by CRISPRi gRNA
|
gRNA sequence (without tag at 3' end)
|
Locus targeted (eg. beginning of gene, putative promoter, -35 region)
|
Target template or nontemplate strand
|
red
|
Ethanol
|
ack (indirectly, pta)
|
GTTTTTTTAGCCACGTATCAATTAT
|
promoter region of ack
|
Nontemplate strand
|
orange
|
Ethanol
|
ldhA
|
ATTCAACATCACTGGAGAAAGTCTT
|
promoter
|
template
|
blue
|
Ethanol
|
ackA
|
TTTTTAGCCACGTATCAATTAT
|
promoter region of ackA, starting at -32
|
nontemplate
|
Team
|
Ethanol (E) or Acetate (A)
|
Gene targeted by CRISPRi gRNA
|
gRNA sequence (without tag at 3' end)
|
Locus targeted (eg. beginning of gene, putative promoter, -35 region)
|
Target template or nontemplate strand
|
Instructor
|
Acetate
|
aceEF
|
GGAATAACCCatgtcagaacgtttc
|
5' UTR region and beginning of coding region
|
Template
|
Instructor
|
Acetate
|
adhE (1)
|
GGAGAGCATTatggctgttactaatg
|
5' UTR region and beginning of coding region
|
Template
|
Instructor
|
Acetate
|
adhE (2)
|
AATGCTCTCCTGATAATGTTAAAC
|
Promoter and 5' UTR region right before coding region
|
Nontemplate
|