ptdTHC
(Plasmid #112617)

• Purpose: promoterless expression plasmid driving multicistronic cassette H2BtdTomato_p2A_HygroR_p2A_CreERt2
• Depositing Lab: Stefan Heller
• Publication: Hartman et al Dev Biol. 2018 Sep 1. pii: S0012-1606(18)30499-8. doi: 10.1016/j.ydbio.2018.08.013.

Backbone

• Vector backbone: promoterless backbone from pd2EGFP-1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3110
• Total vector size (bp): 8840
• Vector type: Mammalian Expression ; Promoterless vector, ready for promoter for mammalian expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: H2B-tdTomato
• Alt name: HIST1H2BB, H2B.1, H2B/f, H2BFF, MGC119804
• Alt name: tdTomato
• Species: H. sapiens (human); Anthozoa
• Insert Size (bp): 1812
• GenBank ID: NM_021058
• Entrez Gene: H2BC3 (a.k.a. H2B.1, H2B/f, H2BFF, HIST1H2BB)
• Promoter: No Promoter
• Tag / Fusion Protein:
  • tdTomato (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: AAAGAATTCATGAGGCCGGCC
• 3′ sequencing primer: TGAAGTTAGTAGCTCCGCTTCC

Gene/Insert 2

• Gene/Insert name: Hygrophosphotransferase
• Alt name: HygroR
• Alt name: hygromycin-B
• Species: Coccidioides posadasii C735 delta SOWgp
• Insert Size (bp): 1023
• GenBank ID: XM_003071606
• Promoter: No Promoter

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: atgaaaaagcctgaactcaccgc
• 3′ sequencing primer: ttcctttgccctcggacga

Gene/Insert 3

• Gene/Insert name: Cre-ERT2 fusion protein
• Species: Bacteriophage P1
• Insert Size (bp): 1983
• Promoter: No Promoter

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ATGTCCAATTTACTGACCGTACA
• 3′ sequencing primer: TCAAGCTGTGGCAGGGAAACC

Resource Information

• Supplemental Documents:
  • ptdTHC.gbk
• A portion of this plasmid was derived from a plasmid made by: H2B was cloned from Addgene plasmid 26750, tdTomato was cloned from Addgene plasmid 26123, HygroR was cloned from Addgene plasmid 28226, CreERT2 and Rabbit betaglobin PolyA were cloned from Addgene plasmid 14797

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Promoterless multicistronic reporter plasmid containing H2BtdTomato, HygroR, and CreERT2 in a single ORF with two P2A sequences.