Table 3
Characteristics of modular vectors for up to 12 gRNAs
Vector
Golden Gate
overlaps
gRNA
promoter
Cas
promoter Cas
PAM
E. coli
Marker
Plant
Marker Reference
PCR template
vectors:
pEN-2xAtU6
—
AtU6–26 —
—
—
Km
—
[10]
Entry vectors:
pGG-AAtU6ccdB-B
AB
AtU6–26 —
—
Amp
—
[10]
pGG-BAtU6ccdB-C
BC
AtU6–26 —
—
Amp
—
[10]
pGG-CAtU6ccdB-D
CD
AtU6–26 —
—
Amp
—
[10]
pGG-DAtU6ccdB-E
DE
AtU6–26 —
—
Amp
—
[10]
pGG-EAtU6ccdB-F
EF
AtU6–26 —
—
Amp
—
[10]
pGG-FAtU6ccdB-G
FG
AtU6–26 —
—
Amp
—
[10]
pGG-BlinkerII-G
BG
—
—
—
Gm
—
[10]
pGG-ClinkerII-G
CG
—
—
—
Gm
—
[18]
pGG-DlinkerII-G
DG
—
—
—
Gm
—
[10]
pGG-ElinkerII-G
EG
—
—
—
Gm
—
[18]
pGG-FlinkerII-G
FG
—
—
—
Gm
—
[10]
Binary vectors:
pFASTGK
AtCas9AccdB-G
AG
—
PcUbi4–2 SpCas9
a NGG Sp
Km,
green
seed
—
pFASTRK
AtCas9AccdB-G
AG
—
PcUbi4–2 SpCas9
a NGG Sp
Km, red
seed
—
Abbreviations: gRNA, guide RNA; PAM, protospacer adjacent motif; Km, kanamycin; Amp, ampicillin; Gm, gentamycin; PPT, phosphinothricin; Sp, spectinomycin; Ref., reference
a
Arabidopsis thaliana codon-optimized Streptococcus pyogenes Cas9
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Gwen Swinnen et al.