pGGY001
(Plasmid #48866)

• Purpose: (Empty Backbone) Empty destination vector for creating a GreenGate plant transformation plasmid with the plant resistance cassette at the T-DNA right border.
• Depositing Lab: Jan Lohmann
• Publication: Lampropoulos et al PLoS One. 2013 Dec 20;8(12):e83043. doi: 10.1371/journal.pone.0083043.

Backbone

• Vector backbone: pGreenII
• Backbone size (bp): 2492
• Vector type: Golden Gate compatible plant transformation vector

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Gentamicin, 25 & 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DB3.1
• Growth instructions: This plasmid should be grown at reduced gentamycin concentration (5 µg/mL).
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: None

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid is designed as destination vector for Golden Gate cloning using the BsaI enzyme. The 5' BsaI overhang can be exchanged using KpnI and EcoRI. The 3' BsaI overhang can be exchanged using XhoI and XbaI. There is also a a NotI-PacI-SgsI-SfaAI-KpnI MCS in front of the A-overhang and an XbaI-PaeI MCS after the G-overhang.

Please note that a transposon insertion is present in the chloramphenicol resistance promoter. This insertion does not interfere with plasmid function. There is an additional BsaI site at bp# 1889-1894 which is irrelevant for GreenGate reactions with this plasmid.

Plasmid Features (listed as bp in full plasmid sequence):
BsaI site #1 = 640-650bp
transposon integration = 747-2077bp
BsaI site #2 = 3414-3419bp