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Purpose(Empty Backbone) Suicide vector for allelic replacement in Bacteroides, tetracycline selection and rhamnose-inducible ss-Bfe1 counterselection
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 126620 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonenew backbone
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Modifications to backboneNEBuilder assembly reaction to fuse: a) rhamnose promoter, amplified from gene BT_3763 from B. thetaiotaomicron VPI-5482 (this promoter drives ss-bfe1 expression) b) ss-bfe1 gene and multiple cloning site, amplified from pLGB13, using oLGB110 and oLGB120 c) tetracycline resistance casette, amplified from vector pYT646b (Tang and Malamy, 2000), using oLGB121 and oLGB122 d) origin of transfer and replication, amplified from pLGB13 e) RhaR cassette (BT_3768), amplified from B.thetaiotaomicron VPI-5482
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Vector typeallellic exchange in bacteria
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Selectable markerstetracycline (Bacteroidales)
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)S17-lambda pir
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Copy numberLow Copy
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pLGB30 was a gift from Laurie Comstock (Addgene plasmid # 126620 ; http://n2t.net/addgene:126620 ; RRID:Addgene_126620) -
For your References section:
Streamlined Genetic Manipulation of Diverse Bacteroides and Parabacteroides Isolates from the Human Gut Microbiota. Garcia-Bayona L, Comstock LE. mBio. 2019 Aug 13;10(4). pii: mBio.01762-19. doi: 10.1128/mBio.01762-19. 10.1128/mBio.01762-19 PubMed 31409684