Table 1 Plasmids, primers and oligo used in the current study

pCas

Kanamycin resistance

Temperature sensitive: cultured at 30 °C, Size: 12,545 bp

[22], Addgene plasmid #62,226

pTargetF

Spectinomycin resistance, Size: 2117 bp

[22], Addgene plasmid #62,226

pTargetF-OmpF

Modified pTargetF; 20 bp change of gRNA

This study

pUC57-ompF-MPER

Plasmid vector containing MPER surrounded with 300 bp (from each side) homologous to ompF sequences.

GenScript

Primers

pTF_ompF107-F

CGCGTCGTTTTAGAGCTAGAAATAGC (Tm¹ = 66.5^oC)

Insertion of gRNA into pTargetF

This study

pTF_ompF107-R

TTTTGTTACCAGTTACTAGTATTATACCTAGGACTGAGTC (Tm¹ = 66.5^oC)

Insertion of gRNA into pTargetF

This study

118 F Pre-OmpF forward primer)

GTAGCACTTTCACGGTAGCG (Tm¹ = 60 ºC)

Binds to the plus strand upstream of ompF

This study

105R2 (OmpF reverse primer)

GTAGCTGATAGAACCGCCAACAC (Tm¹ = 60 ºC)

Binds to the minus strand of ompF, downstream the insertion

This study

108 F (pTargetF forward primer)

TTTCCTGCGTTATCCCCTGA (Tm¹ = 60ºC)

Binds to the plus strand of pTargetF, upstream the sgRNA

This study

108R (pTargetF reverse primer)

CGACGCGTTTTGTTACCAGT (Tm1 = 60 ºC)

Binds to the minus strand of pTargetF-OmpF and is specific for the OmpF-gRNA

This study

109 F (OmpF forward primer)

AACAGTTACGGTGGCAATGG (Tm1 = 60 ºC)

Binds to the plus strand of ompF, upstream the insertion

This study

109R (MPER reverse primer)

CCAAAGACTTGCCCACTTGT (Tm1 = 60 ºC)

Binds to the minus stand of MPER

This study