pANDA- vector map

BamH I
EcoR I
Site D
Site C

Site E
Site A

Site B

Sac I
Hind III, Pst I,
Pst I Pst I EcoR I EcoR I Sal I
RB LB
gus
NPT II Ubq pro. attR Ubq pro. bar Nt HPT
attR
linker Nt

1.9 Kbp 1.7 Kbp 1 Kbp 1.7 Kbp 1.9 Kbp 0.7 Kbp
0.3 Kbp

Ubq pro. attR

+1 +84 +85
intron
Ubq pro. attR1 CmR ccdB attR2

0.9 Kbp 1 Kbp 1.7 Kbp

LB: left border

RB: right border
NPT II: Kanamycin resistance gene
HPT: Hygromycin resistance gene
Ubq pro.: Maize ubiquitin1 promoter + 1st intron & splicing acceptor site
attR: LR clonase recombination cassette (Invitrogen, Cat. No. 11828-019, rfA)
attR1 & attR2: LR clonase recombination sites
CmR: Chloramphenicol resistance gene
ccdB: ccd B gene
Nt: NOS terminator
bar: Bialaphos resistance gene
Vector size: about 20 Kbp
Back bone: pBI101
Host E. coli strain: DB 3.1
Kpn I is unique restriction enzyme site, others are not unique.

Site A
RB---BamH I, Sma I, Kpn I, Apa I, Xho I ---LB
Site B
RB---Cla I, Hind III, EcoR V ---LB
Site C
RB---EcoR I, Pst I, Sma I, BamH I, Xho I, Not I, EcoR V, Pst I, EcoR I ---LB
Site D
RB---EcoR I, Spe I, BamH I, Sac I ---LB
Site E
RB---EcoR I, EcoR V, Hind III ---LB