Table 2 Strains, plasmids, and oligonucleotides used in this study

BL21-Gold (DE3)

E. coli B F^– ompT hsdS(rB^– mB^–) dcm⁺ Tet^R gal λ(DE3) endA Hte

Agilent technologies

XL1 Blue

recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F’ proAB lacIqZΔM15 Tn10 (Tetr)]

Agilent technologies

Cupriavidus necator H16

Wild type, PHA producing

ATCC 17699

Plasmids

pLG575

pACYC184 derivative, HlyBD, p15A origin, Cm^R

[47]

pBHR68

pBluescript SK-, phbCAB genes from R. eutropha

[16]

pSB1AK3

High copy BioBrick vector, pMB1 origin, Amp^R and Kan^R

[49]

pSB1A3

High copy BioBrick vector, pMB1 origin, Amp^R

[49]

pSB3K3

Medium copy BioBrick standard vector, p15A origin, Kan^R

[49]

pCMEL1

phaP1, C-terminal BioFusion with HlyA signal peptide, Lac promoter (BBa_R0010), RBS(BBa_B0034), in pSB1A3

This study

pCMEL2

phaP1, C-terminal BioFusion with HlyA signal peptide, Lac promoter (BBa_R0010), RBS(BBa_B0034), in pSB3K3

This study

pCMEL3

phaP1, C-terminal BioFusion with HlyA signal peptide, Lac promoter (BBa_R0010), RBS(BBa_B0034), in pBHR68

This study

Oligonucleotides

PhaP1FOR

5′- gaattcgcggccgcttctagaatgatcctcaccccggaaca-3′

This study

PhaP1REV

5′- ctgcagcggccgctactagttcaggcagccgtcgtcttct-3′

This study

g114t

5′- cgtcgagctgaaccttcaggtcgtcaagact-3′

This study

g114t_antisense

5′-agtcttgacgacctgaaggttcagctcgacg-3′

This study