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Distribution host: Escherichia coli JM101

Size (kb): 13.1000003814697300

DESCRIPTION OF VECTOR:
Intact vector size: 13.100
Type of vector: cosmid
Cloning sites: BamHI HindIII NheI
Polylinker sites:
Cloning capacity: 30-42 kb
Construction: Lawrist4, pYUB12
Host range: Mycobacterium sp.; Escherichia coli
Features (with orientation and position when available):
marker(s): ampR, kanR
replicon: lambda, pMB1, pAL5000
terminator: trpA, rrnC, fd

Vector: pMSC1 (cosmid)

Construction: Lawrist4, pYUB12

Marker(s):ampR,kanR

Construct size (kb): 13.10000038146973

Features: marker(s): ampR, kanR
replicon: lambda, pMB1, pAL5000
terminator: trpA, rrnC, fd

shuttle vector

vector for constructing genomic libraries

Restriction digests of the clone give the following sizes (kb): EcoRV-8.2, 5.0; HindIII--13.5; BamHI--13.5; NheI--13.5; EcoRI--6.5, 2.5, 1.3.

The pAL5000 ori permits replication in Mycobacterium smegmatis and M. bovis BCG.

The vector should be selected using ampicillin and kanamycin in E. coli, and kanamycin in Mycobacterium spp.

IMPORTANT: To prevent amplification of a rearranged and/or deleted cosmid, we recommend streaking on LB + amp plates at 30C and picking small colonies for liquid culture.

A cosmid shuttle vector useful for genomic DNA library construction.

The ScaI site between the cos sites can be used to linearize the plasmid. After packaging, 10.3 kb of vector is retained, permitting an insert range of 30 - 42 kb. Sequences hybridizing to pBR322, including ampR, are removed.

The order of the major features in this plasmid is: cos - pMB1 ori - ampR/ScaI - cos - trpA terminator - XbaI - HindIII - fd terminator - EcoRV - pAL5000 ori - BamHI - rrnC terminator - EcoRV - lambda ori - NheI - kanR.

Temperature: 30.0°C