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Recombinant colonies pCB2004-Cs F3′5′H and control pCB2004 vectors were selected on kanamycin plates and validated by restriction enzyme digestion, followed by transformation into EHA105 by electroporation at 2500 V for about 5.5 ms.

Four independent shRNA viral vectors, targeting different Atf4 mRNA sequences, were used and one shRNA vector was selected on the basis of the highest silencing efficacy.

After recombination, the resulting pBS-attR3-ccdB-CmR-attR5 vector was selected on ampicillin and chloramphenicol LB agar DB3.1 bacteria.

Transformants containing modified ClosTron vectors were selected from E. coli strain DH10B based on chloramphenicol resistance, and plasmid DNA was isolated using a plasmid minipreparation kit (Fermentas Inc., Glen Burnie, MD).

T1 seeds of amiRNA and empty-vector lines were selected on 40 mg/L hygromycin.

Seeds obtained from transgenic and vacant vector lines were selected on MS medium with 50 mg/L hygromycin.

The amplified product was co-transformed into yeast (ura3- 1) with the gapped vector, and transformants were selected on -URA.

Depending on the vector used, transformants were selected on MS medium [ 50] containing 50 mg/L hygromycin or 100 mg/L kanamycin, 500 mg/L cefotaxime, 1 mg/L BAP and 0.1 mg/L NAA.

*Control TAR cloning experiment was carried out with the same TAR vector but transformants were selected on SD medium lacking 5-FOA Because the human genome contains approximately 250,000 copies of alphoid DNA monomer, it is possible that alphoid DNA could be cloned into an ARS-containing vector even without counter-selection.

Primary transgenic lines for each amiRNA vector were selected based on their phenotype.

The NiCo21 DE3) were individually transformed with pACYC-RIL, pRARE2, and pLysSRARE2 vectors and the transformants were selected on LB agar plates containing 25 μg mL-1 Chloramphenicol.