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Vector Backbone Description: Backbone Size:4800; Vector Backbone:mTopaz-pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Contains 6x-Histidine tag for protein purification. Excitation = 514; Emission = 527
Proper citation: RRID:Addgene_54841 Copy
Vector Backbone Description: Backbone Size:4800; Vector Backbone:mVenus-pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11753368
Comments: Contains 6x-Histidine tag for protein purification. Excitation = 515; Emission = 528
Proper citation: RRID:Addgene_54845 Copy
Genetic Insert: mTurquoise2
Vector Backbone Description: Backbone Size:4800; Vector Backbone:mTurquoise2-pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Contains 6x-Histidine tag for protein purification. Excitation = 434; Emission = 474
Proper citation: RRID:Addgene_54844 Copy
Vector Backbone Description: Backbone Size:4800; Vector Backbone:rsEmerald-pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Contains 6x-Histidine tag for protein purification. Excitation = 491; Emission = 508
Proper citation: RRID:Addgene_54850 Copy
Vector Backbone Description: Backbone Size:4800; Vector Backbone:mOrange-pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: Contains 6x-Histidine tag for protein purification. Excitation = 548; Emission = 562
Proper citation: RRID:Addgene_54751 Copy
Vector Backbone Description: Backbone Size:4800; Vector Backbone:TurboRFP-pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17572680
Comments: Contains 6x-Histidine tag for protein purification. Excitation = 553; Emission = 574
Proper citation: RRID:Addgene_54858 Copy
Vector Backbone Description: Backbone Size:4800; Vector Backbone:tdTurboRFP-pBAD; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17572680
Comments: Contains 6x-Histidine tag for protein purification. Excitation = 553; Emission = 576
Proper citation: RRID:Addgene_54857 Copy
Vector Backbone Description: Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. All 2-series vectors work as single-expression vectors, as well as transfer vectors for our polycistronic system.
The LIC cloning site is flanked by 5 pairs of restriction sites, so that your gene can easily be subcloned into our polycistronic destination vectors (2D, 2E, or 2Z).
2RRT has a TEV-cleavable N-terminal StrepII-StrepII tag.
To clone into this vector, add LIC v1 tags to the 5' end of your PCR primers.
Forward - 5'-TACTTCCAATCCAATGCA-3'
Reverse - 5'-TTATCCACTTCCAATGTTATTA-3'
Linearize the plasmid with SspI and gel purify.
When digesting the DNA with T4 polymerase, use dCTP for insert and dGTP for vector.
Proper citation: RRID:Addgene_55215 Copy
Vector Backbone Description: Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. All 2-series vectors work as single-expression vectors, as well as transfer vectors for our polycistronic system.
The LIC cloning site is flanked by 5 pairs of restriction sites, so that your gene can easily be subcloned into our polycistronic destination vectors (2D, 2E, or 2Z).
2-HpI-69 has a TEV-cleavable N-terminal His6-High pI protein fusion tag. The high pI can improve solubility of your target protein.
To clone into this vector, add LIC v1 tags to the 5' end of your PCR primers.
Forward - 5'-TACTTCCAATCCAATGCA-3'
Reverse - 5'-TTATCCACTTCCAATGTTATTA-3'
Linearize the plasmid with SspI and gel purify.
When digesting the DNA with T4 polymerase, use dCTP for insert and dGTP for vector.
Proper citation: RRID:Addgene_55212 Copy
Vector Backbone Description: Vector Backbone:pET; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: This plasmid is an empty vector. Your gene can be inserted with a LIC cloning protocol. All 2-series vectors work as single-expression vectors, as well as transfer vectors for our polycistronic system.
The LIC cloning site is flanked by 5 pairs of restriction sites, so that your gene can easily be subcloned into our polycistronic destination vectors (2D, 2E, or 2Z).
2-HpI-68 has a TEV-cleavable N-terminal His6-High pI protein fusion tag. The high pI can improve solubility of your target protein.To clone into this vector, add LIC v1 tags to the 5' end of your PCR primers.
Forward - 5'-TACTTCCAATCCAATGCA-3'
Reverse - 5'-TTATCCACTTCCAATGTTATTA-3'
Linearize the plasmid with SspI and gel purify.
When digesting the DNA with T4 polymerase, use dCTP for insert and dGTP for vector.
Proper citation: RRID:Addgene_55211 Copy
Vector Backbone Description: Vector Backbone:pFastBac; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28668116
Comments: In order to increase the efficiency of subcloning with the Series-11 Macrobac plasmids we developed a new strategy the uses LIC (Ligation Independent Cloning). The problem with the Series-11 ligation mediated subcloning was that when the plasmid size approached or exceeded 20 kb we had to screen many colonies to find a positive. We developed a strategy to use LIC for subcloning. Because this method uses no ligase, the cloning background associated with re-ligation of the empty plasmid are eliminated.
438-B has a TEV cleavable His6 at the N-terminus.
The 438-B vector use the LICv1 Forward and Reverse primers.
LICv1Forward - 5'-TACTTCCAATCCAATGCA-3'
LICv1 Reverse - 5'-TTATCCACTTCCAATGTTATTA-3'
As the target plasmid size gets >20kb you may have to increase the amount of DNA you anneal and/or transform. We have found this method gives no background colonies at all and 100% of the colonies we pick are positive. The cells we transform into are XL1Blues with a competency ~6x10^7cfu/ul.
For more information, please see our website: http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_55219 Copy
Vector Backbone Description: Vector Backbone:pFastBac; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28668116
Comments: In order to increase the efficiency of subcloning with the Series-11 Macrobac plasmids we developed a new strategy the uses LIC (Ligation Independent Cloning). The problem with the Series-11 ligation mediated subcloning was that when the plasmid size approached or exceeded 20 kb we had to screen many colonies to find a positive. We developed a strategy to use LIC for subcloning. Because this method uses no ligase, the cloning background associated with re-ligation of the empty plasmid are eliminated.
438-A is an untagged pFastBac LIC Subcloning vector.
The 438 vectors use the LICvBac Forward primer and the LICv1 Reverse primer.
LICvBacF - 5'-TACTTCCAATCCAATCG-3' (Note: ATG must be added to ORF)
LICv1 Reverse - 5'-TTATCCACTTCCAATGTTATTA-3'
As the target plasmid size gets >20kb you may have to increase the amount of DNA you anneal and/or transform. We have found this method gives no background colonies at all and 100% of the colonies we pick are positive. The cells we transform into are XL1Blues with a competency ~6x10^7cfu/ul.
For more information, please see our website: http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_55218 Copy
Vector Backbone Description: Backbone Marker:Life Technologies; Backbone Size:6383; Vector Backbone:pFastBac; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23410102
Comments: C-terminal tags are separated by a stop codon. Depending on the LP2 primer used in SLIC, the desired C-terminal tag(s) can be fused to the target gene.
Use the one of the following linearization primers pairs for SLIC cloning. Choose one LP1 forward vector primer and one LP2 reverse vector primer:
LP1 forward vector primer:
SLIC Primer (3C site) 5' GGGCCCCTGGAACAGAACTTCCAG 3'
LP2 - select an LP2 primer below depending on which C-terminal tags you want to include fused to your gene of interest:
none SLIC Primer 5' CGCCATTAACCTGATGTTCTGGGG 3'
10His- SLIC Primer 5`GAGCATCATCATCATCACCAC 3'
StrepOne - SLIC Primer 5`AGCGCTTGGAGCCACCCGCAG 3'
S-Tag - SLIC Primer 5`AAAGAAACCGCTGCTGCTAAATTCG 3'
HPC4 - SLIC Primer 5`GAGGACCAGGTGGACCCCCGG 3'
Æ54CPD - SLIC Primer 5`GGCAGCGGCAAGATCCTGCAC 3'
Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected.
Proper citation: RRID:Addgene_55185 Copy
Vector Backbone Description: Backbone Marker:Life Technologies; Backbone Size:6063; Vector Backbone:pFastBac; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:23410102
Comments: C-terminal tags are separated by a stop codon. Depending on the LP2 primer used in SLIC, the desired C-terminal tag(s) can be fused to the target gene
Use the one of the following linearization primers pairs for SLIC cloning. Choose one LP1 forward vector primer and one LP2 reverse vector primer:
LP1 forward vector primer:
SLIC Primer (3C site) 5' GGGCCCCTGGAACAGAACTTCCAG 3'
LP2 - select an LP2 primer below depending on which C-terminal tags you want to include fused to your gene of interest:
none SLIC Primer 5' CGCCATTAACCTGATGTTCTGGGG 3'
10His- SLIC Primer 5`GAGCATCATCATCATCACCAC 3'
StrepOne - SLIC Primer 5`AGCGCTTGGAGCCACCCGCAG 3'
S-Tag - SLIC Primer 5`AAAGAAACCGCTGCTGCTAAATTCG 3'
HPC4 - SLIC Primer 5`GAGGACCAGGTGGACCCCCGG 3'
Æ54CPD - SLIC Primer 5`GGCAGCGGCAAGATCCTGCAC 3'
Test each preparation of the plasmid by transforming it into non-resistant cells (ex. DH5a) to ensure negative selection by ccdB kills all the transformed cells as expected.
Proper citation: RRID:Addgene_55184 Copy
Vector Backbone Description: Vector Backbone:pFastBac; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28668116
Comments: In order to increase the efficiency of subcloning with the Series-11 Macrobac plasmids we developed a new strategy the uses LIC (Ligation Independent Cloning). The problem with the Series-11 ligation mediated subcloning was that when the plasmid size approached or exceeded 20 kb we had to screen many colonies to find a positive. We developed a strategy to use LIC for subcloning. Because this method uses no ligase, the cloning background associated with re-ligation of the empty plasmid are eliminated.
438-SNAP-V3 has a TEV cleavable TwinStrepII-Prescission-SNAPf on the C-terminus.
The 438-SNAP-V3 vector use the LICv3 Forward and Reverse primers.
LICv3 Forward - 5'-TTTAAGAAGGAGATATAGTTC-3'
LICv3 Reverse - 5'-GGATTGGAAGTAGAGGTTCTC-3'
As the target plasmid size gets >20kb you may have to increase the amount of DNA you anneal and/or transform. We have found this method gives no background colonies at all and 100% of the colonies we pick are positive. The cells we transform into are XL1Blues with a competency ~6x10^7cfu/ul.
For more information, please see our website: http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_55223 Copy
Vector Backbone Description: Vector Backbone:pFastBac; Vector Types:Insect Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:28668116
Comments: In order to increase the efficiency of subcloning with the Series-11 Macrobac plasmids we developed a new strategy the uses LIC (Ligation Independent Cloning). The problem with the Series-11 ligation mediated subcloning was that when the plasmid size approached or exceeded 20 kb we had to screen many colonies to find a positive. We developed a strategy to use LIC for subcloning. Because this method uses no ligase, the cloning background associated with re-ligation of the empty plasmid are eliminated.
438-Rgfp has a TEV cleavable StrepII and msfGFP at the N-terminus.
The 438-Rgfp vector use the LICv1 Forward and Reverse primers.
LICv1Forward - 5'-TACTTCCAATCCAATGCA-3'
LICv1 Reverse - 5'-TTATCCACTTCCAATGTTATTA-3'
As the target plasmid size gets >20kb you may have to increase the amount of DNA you anneal and/or transform. We have found this method gives no background colonies at all and 100% of the colonies we pick are positive. The cells we transform into are XL1Blues with a competency ~6x10^7cfu/ul.
For more information, please see our website: http://qb3.berkeley.edu/qb3/macrolab/
Proper citation: RRID:Addgene_55221 Copy
Species: Synthetic
Genetic Insert: 3xFVN-Tel-TAL
Vector Backbone Description: Backbone Marker:Hodaka Fujii; Backbone Size:5758; Vector Backbone:pMXs-puro; Vector Types:Mammalian Expression, Retroviral, TALEN; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24201379
Proper citation: RRID:Addgene_63590 Copy
Species: S. pyogenes
Genetic Insert: humanized S. pyogenes dCas9
Vector Backbone Description: Vector Backbone:pUC ori vector; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:25997509
Proper citation: RRID:Addgene_63596 Copy
Species: S. pyogenes
Genetic Insert: humanized S. pyogenes dCas9
Vector Backbone Description: Vector Backbone:pUC ori vector; Vector Types:Mammalian Expression, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:25997509
Proper citation: RRID:Addgene_63597 Copy
Species: Synthetic
Genetic Insert: 3xFVN-Tel-TAL
Vector Backbone Description: Backbone Marker:Hodaka Fujii; Backbone Size:6271; Vector Backbone:pMXs-neo; Vector Types:Mammalian Expression, Retroviral, TALEN; Bacterial Resistance:Ampicillin
Defining Citation: PMID:24201379
Proper citation: RRID:Addgene_63591 Copy
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