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Purpose(Empty Backbone)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 30122 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepFastBac Dual
- Backbone size (bp) 5306
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Vector typeInsect Expression
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Selectable markersGentamicin
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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameNone
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Tag
/ Fusion Protein
- His6-TEV (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site LIC site (destroyed during cloning)
- 3′ cloning site LIC site (destroyed during cloning)
- 5′ sequencing primer see notes
- 3′ sequencing primer see notes (Common Sequencing Primers)
Resource Information
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Articles Citing this Plasmid
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.
Depositor Comments
This plasmid is a LIC-adapted pFastBac Dual vector. It uses the same PCR primer tags as with most of our other vectors, so one PCR product can be inserted into many different vectors at once.
This vector is a dual expression vector, and your genes can be inserted in any order (check your gene for internal restriction sites, as this may dictate cloning order).
A gene inserted into LIC site 1 will have a His6-TEV tag added to the N-terminus. A gene inserted into LIC site 2 will remain untagged.
LIC site v1:
Add the following tags to your PCR primers:
LicV1 Forward Tag TACTTCCAATCCAATGCA
LicV1 Reverse Tag TTATCCACTTCCAATGTTATTA
Linearize vector with SspI and gel purify. T4-treat vector with dGTP. For the PCR product, T4-treat with dCTP.
LIC site v2:
Add the following tags to your PCR primers:
LicV2 Forward Tag TTTAAGAAGGAGATATAGATC(ATG)
LicV2 Reverse Tag TTATGGAGTTGGGATCTTATTA
Linearize vector with EcoRV and gel purify. T4-treat vector with dCTP. For the PCR product, T4-treat with dGTP.
To sequence LIC site v1, use the following primers:
LicBac dual V1 F cctataactattccggattattcataccgtc
LicBac dual V1 R caggttcagggggaggtgtg
To sequence LIC site v2, use the following primers:
LicBac dual V2 F gtcatagcgcgggttccttcc
LicBac dual V2 R ggagtatacggacctttaattcaaccc
For more information, please see our website:
http://qb3.berkeley.edu/qb3/macrolab/
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:
pFastBac Dual His6 TEV LIC cloning vector (5B) was a gift from Scott Gradia (Addgene plasmid # 30122 ; http://n2t.net/addgene:30122 ; RRID:Addgene_30122)