Split selection landing pad & in-situ assembly protocol & updates

Contact Information:

Zach Stevenson: zstevens at uoregon.edu

Megan Moerdyk-Schauwecker: mmoerdyk at uoregon.edu

Article: https://www.g3journal.org/content/early/2020/08/19/g3.120.401400
Protocol Link: (https://drive.google.com/file/d/1Kg_0jkiSXhNU0EhHxkLUoWWjghOudA70/view?usp=sharing)


Plasmids & Sequence Files- Benchling Links:

Plasmids:
pMS81 (https://benchling.com/s/seq-R4EH8NBa1F1H664YrV7B)
Insertion vector plasmid for split integration in the chromosome II site
Addgene: https://www.addgene.org/154840/

pMS79 (https://benchling.com/s/seq-D33sAg6kZoLrCvr13n8J)
Cas9 & guide all-in-one plasmid targeting the PX696 landing pad
Addgene: https://www.addgene.org/154839/

pZCS16 (https://benchling.com/s/seq-054XEhkchQaadFDv1u7G)
Optional co-marker
Addgene: https://www.addgene.org/154824/

pZCS52 (https://benchling.com/s/seq-chIp5pVybr0toshIMyYG)
sqt-1(e1350)donor homology
Avalible upon request

pZCS23 (https://benchling.com/s/seq-rEXZfb52mc7Gr33eVOXl)
CRE expressing plasmid
Avalible upon request

pMS74 (https://benchling.com/s/seq-CN2GupAnLSrA0g9IKHPb)
Vector for HygR LP insertion, SG5
Addgene: https://www.addgene.org/154838/

pMS4 (https://benchling.com/s/seq-pOsYKSUUVy1ytoDI8t1Y)
ChrII insertion vector with SEC
Addgene: https://www.addgene.org/154837/

Strain:
PX696 (https://benchling.com/s/seq-QKhVdT5ErPJdqrRh8Sw1)
The chromosome II insertion strain sequence
CGC: https://cgc.umn.edu/strain/PX696

To use these files in software like ApE, download the .gb file format
ApE can be downloaded at: https://jorgensen.biology.utah.edu/wayned/ape/

Future strain landing pad strains on Chr. I and III along with SEC vectors will be published here once they are ready for distribution.