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Distribution host: Escherichia coli DH5alpha

Size (kb): 10.1000003814697300

DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pRc/RSV
Intact vector size: 5.100
Type of vector: phagemid
Vector end: HindIII
Vector end: XbaI
Cloning sites: HindIII SpeI BstXI NotI XbaI
Polylinker sites: HindIII KpnI SacI BamHI SpeI XmaIII BstXI EcoRI PstI EcoRV
BstXI NotI XhoI SphI NsiI XbaI
Construction: pUC19, RSV, bovine growth hormone
Host range: vertebrate cells; Escherichia coli
Features (with orientation and position when available):
marker(s): ampR, neoR, G418R
promoter: RSV LTR
replicon: pMB1, M13
terminator: bGH polyadenylation
Cross references:

Vector: pRSVPAP (phagemid)

Promoters: Promoter RSV LTR

Construction: pRc/RSV, pSV2Apap

Marker(s):G418R,ampR,neoR

Construct size (kb): 10.10000038146973

Features: marker(s): ampR, neoR, G418R
promoter: RSV LTR
replicon: pMB1, M13
terminator: bGH polyadenylation
enhancer: RSV LTR

contains sequence alkaline phosphatase, placental (Regan isozyme)

expression vector

reporter construct

shuttle vector

vector permitting production of single-stranded DNA

vector permitting visual detection of activity in colonies

Restriction digests of the clone give the following sizes (kb): EcoRI--6.1, 4.0; HindIII--10.1; BglII--10.1; KpnI--5.0, 2.8, 2.3; XbaI--10.1.

Transient expression experiments should be performed to examine the RSV promoter efficiency in different cell lines. The RSV promoter can be replaced by excising with a BglII/HindIII digest.

Shuttle reporter plasmid permitting visual detection of activity by histochemical staining.

Presence of alkaline phosphatase activity is used to follow cell lineage in culture or in situ.

pRSVlacZII (ATCC 77129), pRSVPAP (ATCC 77130) and pRSVADH (ATCC 77131) provide distinct color-staining reactions (aqua blue, red, and blue-black respectively) to permit simultaneous analysis of multiple lineages.

The order of the major features in pRc/RSV is: RSV LTR - MCS - bovine growth hormone polyadenylation signal - M13 ori - SV40 early promoter - neoR - SV40 polyadenylation signal - pMB1 ori - bla.

Constructed by cloning the HindIII/XbaI fragment from pSVApap into pRc/RSV.

Temperature: 37.0°C