15 files

Supplemental Material for Yang et al., 2021

posted on 29.11.2021, 19:05 by Fang-Jung Yang, Chiao-Nung Chen, Tiffany Chang, Ting-Wei Cheng, Ni-Chen Chang, Chia-Yi Kao, Chih-Chi Lee, Yu-Ching Huang, Jung-Chen Hsu, Jengyi Li, Meiyeh J. Lu, Shih-Peng Chan, John Wang
Supplementary File S1. This file. Supplemental methods and results.
Supplementary File S2. Sequence and annotation for the plasmid used to generate the phiC31 landing pad via MosSCI. SnapGene compatible file.
Supplementary File S3. PhiC31 recombination mediated cassette exchange (RMCE) protocol.
Supplementary File S4. Fasta file containing 7 sequences: attB_retrofitted_Sinv47D10_short (original and partially corrected) and attB_retrofitted_Sinv47D10 (original and partially corrected) BACs as well as the LG II sequences, which carry the antIs31, antIs32, and antIs33 sequences, for BRC0566, BRC0906, and BRC0882, respectively.

Supplementary Figure S1. Construction of pCG150_double_phiC31_attB plasmid.
Supplementary Figure S2. Design of dual color operon reporters.
Supplementary Figure S3. Ladder PCR of single-copy insertions of transgenes.
Supplementary Figure S4. Confirmation that phiC31 expressed from the antIs30 transgene is spliced.
Supplementary Figure S5. phiC31 integrase mediated RMCE using antIs30 in C. elegans.
Supplementary Figure S6. Validation of 33.4 kb BAC integration.
Supplementary Figure S7. Large BAC integration attempt.

Supplementary Table S1. Strains used and generated in this study.
Supplementary Table S2. Information for RMCE and other plasmids used in this study.
Supplementary Table S3. Primers used in this study and associated important sequences.
Supplementary Table S4. PCR assay results for candidate insertion lines derived from the longer BAC, attB_retrofitted_Si47D10


Article title

¬¬phiC31 integrase for recombination mediated single copy insertion and genome manipulation in C. elegans