Addgene - pHAGE hSPC-eGFP-W Plasmid Data

Plasmid 36450: pHAGE hSPC-eGFP-W

Gene/Insert 1
Gene/insert name:
human SP-C promoter
Insert size:
3813
Species:
H. sapiens (human)
Gene/Insert 2
Gene/insert name:
eGFP
Insert size:
719
Entrez Gene:
eGFP (pPRS3a_01)
Vector backbone:
pHAGE
(Search Vector Database)
Backbone manufacturer:
Richard Mulligan
Vector type:
Lentiviral
Backbone size w/o insert (bp):
4874
Cloning information for Gene/Insert 1
Promoter:
human SP-C promoter
Cloning site 5':
Not1
Site destroyed during cloning:
No
Cloning site 3':
BamH1
Site destroyed during cloning:
No
5' sequencing primer:
ACGGACACATATAAGACCCTGGTCACA List of Sequencing Primers
3' sequencing primer:
GGAGCAACATAGTTAAGAATACCAGTCAATCTTTC
Cloning information for Gene/Insert 2
Promoter:
N/A
Cloning site 5':
Nde1
Site destroyed during cloning:
Unknown
Cloning site 3':
Not1
Site destroyed during cloning:
No
5' sequencing primer:
CCATTATCGTTTCAGACCCACCTCC List of Sequencing Primers
3' sequencing primer:
TCGCCGTCGAACTTCACCTCG
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Person or lab that originally
cloned the gene/insert:
human 3.7 kb SP-C promoter obtained from Whitsett Lab (Whitsett 3.7SPC/SV40 plasmid).
Sequence: View sequences (3)
Map:
View map
Principal Investigator:
Darrell Kotton
Terms and Licenses:
MTA

Comments:

Cloned h3.7kb SP-C promoter (from Whitsett 3.7SPC/SV40 plasmid)
into pHAGE-CMV-GFP-W as follows:

SV40t removed from SPC plasmid by BamH1-BamH1 cutting, blunting and self-ligtion of blunt ends. Entire hSPC promoter lifted out by cutting Nde1-Not1 and ligated into pHAGE CMV-GFP-W backbone pre-cut Nde1-Not1 to remove most of CMV promoter. Cut this backbone Spe1-Nde1 to remove residual CMV promoter fragment, blunted and ligated
NB: final vector still has Spe1 site at start of promoter.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Efficient derivation of purified lung and thyroid progenitors from embryonic stem cells. Longmire et al (Cell Stem Cell. 2012 Apr 6;10(4):398-411. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 36450" in your Materials and Methods section.