pMyt1GFPicre-Knock-In
(Plasmid #22657)

• Depositing Lab: Lynn Hudson
• Publication: Hudson lab targeting plasmid (unpublished)

Backbone

• Vector backbone: pOSfrtloxP
• Backbone manufacturer: gift from Dr. Randy Thresher
• Backbone size w/o insert (bp): 8272
• Vector type: Targeting vector

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: DH5 alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Myelin Transcription Factor 1
• Alt name: Myt1
• Species: M. musculus (mouse)
• Insert Size (bp): 6302
• Entrez Gene: Myt1 (a.k.a. Nzf2, Nzf2a, Nzf2b, Nztf2)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Multiple (unknown if destroyed)
• 3′ cloning site: Multiple (unknown if destroyed)
• 5′ sequencing primer: n/a

Terms and Licenses

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

Depositor Comments

4899bp of genomic Myt1 sequences with MET of Exon 1 as 5' region of homolgy (NotI/PmeI). Inserted upstream of eGFP/icre/frt/loxP. 1403bp of genomic Myt1 sequence starts at exon 2 and extends past Exon 3 as 3' region of homology (ClaI/NheI) Inserted downstream of eGFP/icre/frt/loxP.

The iCre-PolyA cassette was PCR'd out of the pBlue.iCre (a gift of Dr. Rolf Sprengel) with HindIII/KpnI ends. The iCre cassette was ligated to peGFP-C2 (HindIII/KpnI) (Clontech). The eGFP-iCre-PolyA cassette was PCR'd out with SalI ends. This eGFP-icre-PolyA (SalI) cassette was then cloned into pOSfrtloxP (Dr. Randy Thresher) upstream of the frt and loxP sites. 4886bp of 5' Mu genomic Myt1 DNA was inserted upstream of eGFP and the 1404 bp 3' Mu Genomic DNA was inserted downstream NEO gene. The entire pMyt1GFPicre-Knock-In has been sequenced and verified.