pHSS6
(Plasmid
#37060)
-
Purpose(Empty Backbone)
-
Depositing Labs
-
Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
---|---|---|---|---|---|
Plasmid | 37060 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
-
Vector backbonepMB8
- Backbone size (bp) 2320
-
Modifications to backboneThis Tn3-free vector was designed to carry a minimal amount of nonessential information to favor transposition into cloned DNA sequences rather than into the vector. The important part of the vector is the polylinker surrounded by restriction endonuclease Not I sites. Since Not I has an 8-bp recognition sequence (5'-GCGGCCGC) it would be predicted to cut once every 16 kilobases (kb) in a random DNA sequence. In an A-T-rich organism like yeast, the enzyme has a probability of cutting every 390 kb (assuming 60% AT). Therefore, DNA fragments that have been cloned between the Not I sites have a high probability of being excised intact by Not I. The basic vector used in the procedure is derived from an amplifiable pMB8 replicon (pFH97; Heffron et al., 1978), the kanamycin resistance determinant (Kmr) from Tn5, and a synthetic double-stranded DNA molecule containing recognition sites for the restriction enzymes Not I, BamHI, and EcoRI. The vectors also contain the polylinker from pLink322 (Maniatis et al., 1982) that contains sites for Cla I, HindIII, Xba I, Bgl II, and Pst I. The vector is 2.3 kb in size and contains less than 1000 bp of nonessential DNA sequence.
-
Vector typeCloning vector
Growth in Bacteria
-
Bacterial Resistance(s)Kanamycin, 50 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberUnknown
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer NA
- 3′ sequencing primer Kan-R (Common Sequencing Primers)
Terms and Licenses
-
Academic/Nonprofit Terms
-
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.
-
For your Materials & Methods section:
pHSS6 was a gift from Fred Heffron & Hank Seifert (Addgene plasmid # 37060 ; http://n2t.net/addgene:37060 ; RRID:Addgene_37060) -
For your References section:
Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae. Seifert HS, Chen EY, So M, Heffron F. Proc Natl Acad Sci U S A. 1986 Feb;83(3):735-9. 10.1073/pnas.83.3.735 PubMed 3003748