pBGE2.0
(Plasmid
#165987)
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PurposeMulti-host shuttle vector that can be transferred by conjugation and replicate in S. meliloti, E. coli, S. cerevisiae, and P. tricornutum. Contains S. meliloti pSymB origin and Tet resistance.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 165987 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepPtGE31
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Backbone manufacturerSlattery et al., 2018
- Backbone size w/o insert (bp) 9507
- Total vector size (bp) 18639
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Modifications to backboneAddition of elements for S. meliloti replication and selection.
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Vector typeSynthetic Biology ; Bacterial and Yeast Cloning; Conjugation
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Selectable markersHIS3 ; Chloramphenicol - E. coli, Tetracycline - S. meliloti, Nourseothricin N-acetyltransferase - P. tricornutum
Growth in Bacteria
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Bacterial Resistance(s)Chloramphenicol, 25 μg/mL
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Growth Temperature37°C
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Growth Strain(s)E. coli Epi300
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Growth instructionsPlasmid can be induced to high copy with arabinose.
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Copy numberLow Copy
Gene/Insert 1
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Gene/Insert nameRK2/RP4 origin of transfer
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Alt nameoriT
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Insert Size (bp)771
Gene/Insert 2
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Gene/Insert namerepA1B1C1
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SpeciesSinorhizobium meliloti
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Insert Size (bp)4497
Gene/Insert 3
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Gene/Insert namenourseothricin N-acetyl transferase
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Alt nameNAT
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Insert Size (bp)570
Gene/Insert 4
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Gene/Insert nametetracycline resistance
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Alt nameTetR/A
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Insert Size (bp)2285
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made by(1) Edgell Lab (backbone was cloned from pPtGE30). Reference: Slattery, S. S., Diamond, A., Wang, H., Therrien, J. A., Lant, J. T., Jazey, T., ... & Edgell, D. R. (2018). An expanded plasmid-based genetic toolbox enables Cas9 genome editing and stable maintenance of synthetic pathways in Phaeodactylum tricornutum. ACS synthetic biology, 7(2), 328-338. (2) Jones and Gutterson (Tetracycline resistance gene was cloned from pRK7813). Reference: Jones, J. D., & Gutterson, N. (1987). An efficient mobilizable cosmid vector, pRK7813, and its use in a rapid method for marker exchange in Pseudomonas fluorescens strain HV37a. Gene, 61(3), 299-306.
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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:
pBGE2.0 was a gift from Bogumil Karas (Addgene plasmid # 165987 ; http://n2t.net/addgene:165987 ; RRID:Addgene_165987) -
For your References section:
Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer. Brumwell SL, MacLeod MR, Huang T, Cochrane RR, Meaney RS, Zamani M, Matysiakiewicz O, Dan KN, Janakirama P, Edgell DR, Charles TC, Finan TM, Karas BJ. PLoS One. 2019 Jun 17;14(6):e0206781. doi: 10.1371/journal.pone.0206781. eCollection 2019. 10.1371/journal.pone.0206781 PubMed 31206509