pHAGE hSPC-eGFP-W
(Plasmid #36450)

• Depositing Lab: Darrell Kotton
• Publication: Longmire et al Cell Stem Cell. 2012 Apr 6;10(4):398-411.

Backbone

• Vector backbone: pHAGE
• Backbone manufacturer: Richard Mulligan
• Backbone size w/o insert (bp): 4874
• Total vector size (bp): 10634
• Vector type: Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: human SP-C promoter
• Species: H. sapiens (human)
• Insert Size (bp): 3813
• Promoter: human SP-C promoter

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: Not1 (not destroyed)
• 3′ cloning site: BamH1 (not destroyed)
• 5′ sequencing primer: ACGGACACATATAAGACCCTGGTCACA
• 3′ sequencing primer: GGAGCAACATAGTTAAGAATACCAGTCAATCTTTC

Gene/Insert 2

• Gene/Insert name: eGFP
• Insert Size (bp): 719
• Entrez Gene: eGFP (a.k.a. pPRS3a_01)
• Promoter: N/A

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: Nde1 (unknown if destroyed)
• 3′ cloning site: Not1 (not destroyed)
• 5′ sequencing primer: CCATTATCGTTTCAGACCCACCTCC
• 3′ sequencing primer: TCGCCGTCGAACTTCACCTCG

Resource Information

• A portion of this plasmid was derived from a plasmid made by: human 3.7 kb SP-C promoter obtained from Whitsett Lab (Whitsett 3.7SPC/SV40 plasmid).
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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.