| Vector |
Vector: pAd.CMV.Link.1
Vector type: plasmid
Vector size (kb): 6.668
Cloning sites: EcoRV ClaI HindIII KpnI SalI XbaI XhoI SpeI
Markers: ampR
Features (with orientation and position (nt) when available):
Restriction site: EcoRI, -->, 2
Other: Ad mu 0-1,
Promoter: CMV immediate early enhancer and B-actin promoter, -->, 381-903
Restriction site: NcoI, 681
Restriction site: XbaI, 979
Restriction site: XhoI, 995
Other: Intron, -->
Restriction site: NotI, 1174
MCS: EcoRV…SpeI, --> 1181-1253
Restriction site: NotI, -->, 1834
Enhancer: SV40 PolyA, -->, 1269-1466
Restriction site: XbaI, SalI, 1473-1479
Restriction site: NcoI SmaI, 1834-2287
Other: Ad 9-16 mu, -->
Restiction site: NcoI, 3619
Replicon: pAT153 |
| Comments |
Restriction digests of the clone give the following sizes (kb): EcoRI--6.2, 2.1, 1.6, 1.1; HindIII--10.0, 0.9; BamHI--6.0, 2.5, 1.6, 0.6, 0.2; BglII--10.6, 0.4; PstI--5.1, 2.3, 1.2, 0.8, 0.7, 0.5, 0.4. Contains the full-length cDNA with expression regulated in mammalian cells by a CMV enhancer and a beta-actin promoter. Digestion of the construct with XbaI yields 2 major fragments of 4.3 kb, containing most of the CFTR sequence and the polyadenylation signals, and 6.7 kb, containing the adenovirus and plasmid vector sequences. Distributed in aliquots of 200 ng (40 ng/ul). The order of the major features in this plasmid is: NheI - Ad 0-1 - CMV enhancer - beta-actin promoter - XbaI - insert (contains 1 XbaI site) - SV40 polyadenylation signal - XbaI - Ad 9.2-16.1 - pAT153. |
