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Species: Homo sapiens
Genetic Insert: Keratin
Vector Backbone Description: Backbone Size:5500; Vector Backbone:mCherry-miniSOG; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21483721
Comments: . Excitation = 448 / 473; Emission = 500 / 528
Proper citation: RRID:Addgene_55091 Copy
Species: Mus musculus
Genetic Insert: Talin
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:22751201
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55139 Copy
Species: Mus musculus
Genetic Insert: SSTR3
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55135 Copy
Species: Mus musculus
Genetic Insert: Smoothened1
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55134 Copy
Species: Homo sapiens
Genetic Insert: NM_003380.3
Vector Backbone Description: Backbone Size:5500; Vector Backbone:mCherry-miniSOG; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21483721
Comments: . Excitation = 448 / 473; Emission = 500 / 528
Proper citation: RRID:Addgene_55099 Copy
Species: Homo sapiens
Genetic Insert: Sequestosome1
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55132 Copy
Species: Homo sapiens
Genetic Insert: TBRII
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: TBRII contains the H385N mutation . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55141 Copy
Species: Mus musculus
Genetic Insert: Talin R
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55140 Copy
Species: Homo sapiens
Genetic Insert: TOMM20
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55146 Copy
Species: Homo sapiens
Genetic Insert: TFR
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55144 Copy
Species: Homo sapiens
Genetic Insert: Tensin1
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55143 Copy
Species: Mus musculus
Genetic Insert: VASP
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55153 Copy
Species: Homo sapiens
Genetic Insert: UbiquitinC
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55150 Copy
Species: Mus musculus
Genetic Insert: WASH1
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: WASH = WASH1. Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55162 Copy
Species: Mus musculus
Genetic Insert: WASF2
Vector Backbone Description: Backbone Size:4750; Vector Backbone:mCherry-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: . Excitation = 587; Emission = 610
Proper citation: RRID:Addgene_55161 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
Species: Synthetic
Genetic Insert: His10
Vector Backbone Description: Backbone Marker:iGEM; Backbone Size:2070; Vector Backbone:pSB1C3; Vector Types:Synthetic Biology, Bacillus BioBrick Box; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:24295448
Comments: For sequencing of insert, use the following primers:
fwd: TGCCACCTGACGTCTAAG
rev: ATTACCGCCTTTGAGTGA
Proper citation: RRID:Addgene_55179 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
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