Vector IG Sequence Link :
General : plasmid ds-DNA 5100 BP
Functions : (cloning)
Selection : ()
Copy Number :
Hosts : (E.coli)(vertebrate cells)(E.coli)(D17-C3 helper cell line for transfection)(D17 cell line for infection)
Suppliers : (ATCC)
Misc.Comments : pJD214Hy was constructed from 1.45 kb of SNV DNA ligated into the BamHI/EcoRI (nt 2066 to nt 4361) fragment of pBR322, a pUC12 polylinker, and the hygromycin phosphotransferase B gene (BamHI fragment blunt end-ligated into the HindIII site of the MCS). [2] An intermediate, pJD217SVHy, was constructed from pJD214Hy by deleting a SacI/AvaI fragment (SNV map units 7.747-8.127) and replacing it with an XhoI linker, and inserting a 565 bp NdeI/HindIII fragment from pSV2-neo (SV40 promoter) at the XbaI site. [3] Constructed from pJD217SVHy by inserting a 220 bp BamHI/HindIII fragment (SV40 poly(A) site) into the BamHI site at the 3' end of U5. [3] A replication-defective retroviral vector which is promoter-deficient in the right LTR and cannot express retroviral RNA. Foreign gene expression is regulated by the SV40 early promoter. [3] Removal of SNV map units 7.747 to 8.127 results in deletion of the entire U3 sequence, except for 10 bp at the 5' end containing the attL site. [1] The 5' 10 bp of U3 in the left LTR were deleted, so there is no homology between the two U3 sequences and the risk of recombination is reduced. [1] pJD220SvHy (ATCC 67393) and pRD8 (ATCC 67394) differ in the orientation of the SV40 promoter/hygromycin phosphotransferase B sequences. pRD8 has an additional 3' RNA processing sequence between the left U5 and the hygromycin sequences. [1] Restriction digests of the clone give the following sizes (kb): BamHI--5.0, 0.4; PstI--2.7, 2.3, 0.4; XbaI--5.2. (ATCC staff) Medium is 1227 LB plus ampicillin.
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