Size (kb): 5.3000001907348630
Vector: BNsp-Neo-Alpha (phagemid)
Promoters: Promoter PARP
Construction: BNsp-Neo-T; pBluescript SK+
Construct size (kb): 5.300000190734863
Features: marker(s): ampR, G418R
replicon: pMB1, f1
Restriction digests of the clone give the following sizes (kb): BglIII, EcoRI--3.0, 1.6, 0.8; SacI--2.9, 2.5; EcoRI--5.4; HindIII, BamHI--2.9, 2.5; HindIII--5.4.
Other restriction sites found in the polylinker region of pBluescript SK+ (for example: SpeI, NotI, SacII, EagI, BstXI, ApaII, AccI, XhoI, HindII, etc.) have not yet been tested for their presence in the insert.
This is an unpublished modification of BNsp-Neo-T (ATCC 37802)
Linearizing with MluI before transfection is essential for efficient integration.
Shuttle vector for expressing foreign DNA polycistronicly from the procyclic acidic repetitive protein (PARP) promoter. Integrates at the beta- alpha tubulin tandem array.
The listed cloning sites are derived from the multiple cloning site of pBluescript KS+ and are located 3' to the 3' splice site of the alpha-tubulin sequence.
The alpha-tubulin sequences are located downstream of the PARP promoter and were derived from the immediate upstream of the ATG in the alpha-tubulin gene to the first EcoRI site located in the upstream region.
Lee MG, Van der Ploeg LH. Homologous recombination and stable transfection in the parasitic protozoan Trypanosoma brucei. Science 250: 1583-1587, 1990. PubMed: 2177225
Lex H Van Der Ploeg, personal communication