pHBFD1Bad
(Plasmid #53564)

• Purpose: Produces an active HBFD (carotenoid 4-hydroxy-β-ring 4-dehydrogenase) enzyme in E. coli when expression is induced with arabinose
• Depositing Lab: Francis X Cunningham Jr
• Publication: Cunningham FX et al Plant Cell. 2011 Aug;23(8):3055-69. doi: 10.1105/tpc.111.086827. Epub 2011 Aug 23.

Backbone

• Vector backbone: pBad/His B
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 4092
• Total vector size (bp): 5271
• Modifications to backbone: pHBFD1Bad was made by amplifying the hbfd1 cDNA in pHBFD1 with the oligonucleotide primers HBFD1Nco-N (5′-CACACCATGGCTCCTGTTCTCCTTG-3′) and HBFD1-C (5′-CTGGGCTACATAATGAATAATCCAATC-3′). The first primer served to add an NcoI site at the initiation codon for the hbfd cDNA. The PCR product was blunt cloned in the EcoRV site of pSTBlue1, excised as an NcoI-XhoI fragment (the XhoI site was in the multiple cloning region of pSTBlue1), and then ligated into NcoI + XhoI digested pBAD/His B.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hbfd1 cDNA
• Alt name: AdKC28
• Alt name: carotenoid 4-hydroxy-β-ring 4-dehydrogenase
• Species: Adonis aestivalis
• Insert Size (bp): 1288
• GenBank ID: DQ902556.1
• Promoter: araBAD

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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