BBa_J99000

BBa_J99000 Version 1

Component

Source: http://parts.igem.org/Part:BBa_J99000

Generated By: https://synbiohub.org/public/igem/igem2sbol/1

Created by: David Bikard

Date created: 2009-12-17 12:00:00

Date modified: 2015-05-08 01:08:32

pSWK oriTRP4
[CmR]
oriVR6K
lambda_attP

Details

• Types: DnaRegion
• Roles: Plasmid_Backbone; plasmid_vector
• Sequences: BBa_J99000_sequence (Version 1)

Description

This vector has the conditional origin of replication oriVR6K and can be maintained only in strains expressing the pir gene. It carries the origin of transfer of RP4 and can thus be used in conjugation assays in a donor cell expressing the RP4 conjugation machinery. It also carries the attP recombination site of the lambda phage in order to permit site specific integration at the attB site of E.coli chromosome mediated by the lambda integrase (Valens et al., 2004).

Notes

attP recombination site of the lambda phage was PCR amplified with oligos
5'-TGGAGCTCAAATCAAATAATGATTTTATTT-3' and 5'-GCTGGAGCTCCATGGTACGCGTGCTAGAGGCA-3', and cloned at the SacI site of pSW23T (Demarre et al., 2005). The multiple cloning site was then replaced with the BioBrick strandard restriction sites through inverse PCR with oligos 'GCTTCTAGAGTACTAGTAGCGGCCGCTGCAGGCGGTGGAGCTCAAATCAAATAATG' and 'AGTACTCTAGAAGCGGCCGCGAATTCTATCAAGCTTATCGATACCGTCGACG'

Source

This plasmid was constructed from pSW23T (Demarre et al., 2005)

Sequence Annotations

Sequence Annotation	Location	Component / Role(s)
RP4oriT	1928,2184	feature/misc, sequence_feature
lambda attP	34,450	sequence_feature, feature/misc
oriVR6K	512,938	sequence_feature, feature/misc
cat [CmR]	1012,1671	CDS, feature/protein

Other Properties

• igem#experience: Works
• igem#sampleStatus: Not in stock
• igem#status: Unavailable
• synbiohub#ownedBy: user/james
• synbiohub#ownedBy: user/myers
• synbiohub#topLevel: BBa_J99000/1

Member of these Collections

• iGEM Parts Registry
• igem_plasmid_vectors