Table 1 Plasmids, strains, and primers used in this study

Plasmids

pMD19-Ts

Ap^r, cloning vector

Takara

pFL-XS

Ap^rCm^r, a BioBrick “T” vector used for functional block assembling

[5]

pHM001

Ap^r, pMD19-Ts derivative containing the sll1423 ORF with flanking regions

This study

pHM002

Ap^rKm^r, pMD19-Ts derivative used to knock out sll1423 (ntcA) by insertional inactivation with kanamycin resistance cassette

This study

pHM003

Ap^rKm^r, pMD19-Ts derivative used to delete sll1423 (ntcA) with kanamycin resistance cassette

This study

pHM004

Ap^r, pFL-XS derivative containing the sll1423 ORF

This study

pHM005

Ap^rGm^r, pFL-XS derivative containing Gm^r-P _cpcB -ntcA expression cassette

This study

pHM006

Ap^rGm^r, pFL-XS derivative used to express Gm^r-P _cpcB -ntcA expression cassette at phaAB locus

This study

pHM008

Ap^rGm^r, pMD19-Ts derivative used to inactivate sll1423 with the gentamycin resistance cassette

This study

pHM009

Ap^rGm^r, pFL-XS derivative containing Gm^r-P _cpcB -efe expression cassette

This study

pHM010

Ap^rGm^r, pFL-XS derivative used to express Gm^r-P _cpcB -efe expression cassette at sll1423 locus

This study

pXX55

Ap^r, pFL-XS derivative containing P _cpcB -efe expression cassette

[5]

pXX58

Ap^rGm^r, pFL-XS derivative containing the Gm^r cassette aacC1

[5]

pXX62

Ap^rGm^r, pFL-XS derivative containing Gm^r-P _cpcB cassette

[5]

pKC104

Ap^r, pMD18-Ts derivative containing upstream and downstream fragments of phaAB operon

[26]

pRL446

Ap^rKm^r, plasmid containing the Km^r cassette

Prof. X. Xu

Synechocystis strains

S. PCC 6803

Synechocystis sp. PCC 6803 wild-type

Prof. X. Xu

XX76

slr0168::Sp^r-P _cpcB -efe

[5]

MH013

sll1423::Km^r (Insertional inactivation)

This study

MH015

sll1423::Km^r (Deletion)

This study

MH017

phaAB::Gm^r-P _cpcB -ntcA

This study

MH021

slr0168::Sp^r-P _cpcB -efe/sll1423::Km^r

This study

MH023

slr0168::Sp^r-P _cpcB -efe/phaAB::Gm^r-P _cpcB -ntcA

This study

XX109

slr0168::Sp^r-P _cpcB -efe/sll1981::Km^r-P _cpcB -efe/slr0370::Cm^r-P _cpcB -efe

[5]

MH039

slr0168::Sp^r-P _cpcB -efe/sll1981::Km^r-P _cpcB -efe/slr0370::Cm^r-P _cpcB -efe/sll1423::Gm^r

This study

MH043

slr0168::Sp^r-P _cpcB -efe/sll1981::Km^r-P _cpcB -efe/slr0370::Cm^r-P _cpcB -efe/sll1423::Gm^r-P _cpcB -efe

This study

Primers (5′→3′) ^a

Detection primers for ntcA site (slr1423) in single-copy efe recombinants

ntcA-d1

GTTACTCAGCACAACGGGGTC

ntcA-d2

TTGCAGCCCTTCGCCAGCTGGCACGTTCACGGTAATGGGG

PvuII

kan-1

CCCCATTACCGTGAACGTGCCAGCTGGCGAAGGGCTGCAA

PvuII

kan-2

GCACTGGTCATAGAGGGTGGCAGCTGGCACGACAGGTTTC

PvuII

ntcA-d3

GAAACCTGTCGTGCCAGCTGCCACCCTCTATGACCAGTGC

PvuII

ntcA-d4

TAACTGACCCCGCAGAATGGC

ntcA-x1

ATGGATCAGTCCCTAACCC

ntcA-x2

TTAGGTAAACTGTTGACTGAGAGC

phaAB-3

GCCTTGGGCTAAGTTATTGAGCG

phaAB-4

TAGGATTCTTGCACAGTACCGC

Quantitative real-time PCR

rnpB-f1

TGAGGACAGTGCCACAGAA

rnpB-f2

AATTCCTCAAGCGGTTCCAC

rrn16Sa-f1

CCAACATCTCACGACACGA

rrn16Sa-f2

ACTAGGCGTGGCTTGTATCG

ntcA-f1

CGGCGGAACGGGTTTATT

ntcA-f2

CAATGACAGGTCGGGATGC

Detection primers for ntcA site (slr1423) in multi-copy efe recombinants

ntcA-5

GGCAGTGTGGAGCGCATGTAAT

ntcA-6

AGTAATCACCGTCAACAATACCGC

slr0168-1

ACCTCTCCACGCTGAATTAG

[5]

slr0168-2

TTCCAGGCCACATTGTTGTC

[5]

sll1981-1

GGGCTTCGTTAGGTTGTGTGGC

[5]

sll1981-2

CCGCATGGCCGTTTCCAACTCC

[5]

slr0370-1

GCCGAGGAATACTTAGCCGATG

[5]

slr0370-2

CTGCCCTATGAACCGAATATGG

[5]