pJD220SVHy [pJd 220SvHy] (ATCC® 67393)

Applications: expression vectorintegrating vectorshuttle vector  /  Depositors: Wisconsin Alumni Res. Fndn., HM Temin, Wisconsin Alumni Res. Fndn.

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Designations pJD220SVHy [pJd 220SvHy]
Depositors Wisconsin Alumni Res. Fndn., HM Temin, Wisconsin Alumni Res. Fndn.
Biosafety Level 1
Vector Information
Size (kb): 5.0999999046325680
Vector: pJD220SVHy (plasmid)
Promoters: Promoter SV40 early
Construction: pSV2-neo, pJD214Hy, SV40 promoter
Marker(s):ampR
Construct size (kb): 5.099999904632568
Features: marker(s): ampR
promoter: SV40 early
replicon: pMB1
terminator: SV40
Applications
expression vector
integrating vector
shuttle vector
Comments
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.
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.
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.
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.
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.
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).
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.
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.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Temin HM, Dougherty JP. Promoter-deficient retroviral vector. US Patent 4,980,289 dated Dec 25 1990

Dougherty JP, Temin HM. High mutation rate of a spleen necrosis virus-based retrovirus vector. Mol. Cell. Biol. 6: 4387-4395, 1986. PubMed: 3796606

Dougherty JP, Temin HM. A promoterless retroviral vector indicates that there are sequences in U3 required for 3' RNA processing. Proc. Natl. Acad. Sci. USA 84: 1197-1201, 1987. PubMed: 3029769

Dornburg R, Temin HM. Retroviral vector system for the study of cDNA gene formation. Mol. Cell. Biol. 8: 2328-2334, 1988. PubMed: 2457150

Shipped freeze-dried
Shipping Information Distributed: freeze-dried
Product Sheet
Product Sheet
Product Sheet
Attachments Vector map for 67393