Skip to main content
. 2013 Aug 29;6:125. doi: 10.1186/1754-6834-6-125

Table 2.

Oligonucleotides used in this study

Number Name Sequence (5′ to 3′) Purpose
 
Cloning
 
 
1
XbaI_prk_FW2
TGACATCTAGATGTCACAACAACAAACAATTG
Cloning of PRK into pUDE046.
2
RV1 XhoI prk
TGACATCTAGATGTCACAACAACAAACAATTG
Cloning of PRK into pUDE046.
 
Primers used for in vivo plasmid assembly
 
3
HR-cbbM-FW-65
TTGTAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGGTACAGCTGGAGCTCAGTTTATCATTATC
Rubisco cbbM cassette for plasmids pUDC075, pUDC099, and pUDC100.
4
HR-cbbM-RV-65
GGAATCTGTGTAGTATGCCTGGAATGTCTGCCGTGCCATAGCCATGTATGCTGATATGTCGGTACCGGCCGCAAATTAAAG
Rubisco cbbM cassette for plasmids pUDC075, pUDC099, and pUDC100
5
linker-cbbO2-pRS416
ATCACTCTTACCAGGCTAGGACGACCCTACTCATGTATTGAGATCGACGAGATTTCTAGGCCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGC
Linker fragment for assembly of plasmid pUDC099.
6
linker-cbbM-GroEL
GACATATCAGCATACATGGCTATGGCACGGCAGACATTCCAGGCATACTACACAGATTCCATCACTCTTACCAGGCTAGGACGACCCTACTCATGTATTGAGATCGACGAGATTTCTAGG
Linker fragment for assembly of plasmid pUDC100.
 
Primers used for in vivo integration assembly
 
7
FW pTDH3- HR-CAN1up
GTTGGATCCAGTTTTTAATCTGTCGTCAATCGAAAGTTTATTTCAGAGTTCTTCAGACTTCTTAACTCCTGTAAAAACAAAAAAAAAAAAAGGCATAGCAAGCTGGAGCTCAGTTTATC
1st cloning expression cassette linker fragment between CAN1 upstream and PRK expression cassette (IMI229), and CAN1up-linker and KlLEU2 expression cassette (IMI232).
8
RV linker-iHR2B
AGATATACTGCAAAGTCCGGAGCAACAGTCGTATAACTCGAGCAGCCCTCTACTTTGTTGTTGCGCTAAGAGAATGGACC
1st cloning fragment: linker fragment between CAN1up-linker and PRK expression cassette (IMI229).
9
RV linker-iHR6
GCTATGACCATGATTACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAAAGCTGGTTGCGCTAAGAGAATGGACC
1st cloning fragment: linker fragment between CAN1up-linker and KlLEU2 expression cassette (IMI232).
10
FW pGAL1-prk HR2B
CAACAAAGTAGAGGGCTGCTCGAGTTATACGACTGTTGCTCCGGACTTTGCAGTATATCTGCTGGAGCTCTAGTACGGATT
2nd cloning fragment: GAL1p-PRK-CYC1t expression cassette (IMI229) from pUDE046.
11
RV CYC1t-prk HR2
GGAATCTGTGTAGTATGCCTGGAATGTCTGCCGTGCCATAGCCATGTATGCTGATATGTCGTACCGGCCGCAAATTAAAG
2nd cloning fragment: GAL1p-PRK-CYC1t expression cassette (IMI229) from pUDE046.
12
FW HR2-cbbQ2-HR3
GACATATCAGCATACATGGCTATGG
3rd cloning fragment: PGI1p-cbbQ2-TEF2t cassette (IMI229).
13
RV HR2-cbbQ2-HR3
GGACACGCTTGACAGAATGTCAAAGG
3rd cloning fragment: PGI1p-cbbQ2-TEF2t cassette (IMI229).
14
FW HR3-cbbO2-HR4
CGTCCGATATGATCTGATTGG
4th cloning fragment: PGK1p-cbbO2-ADH1t cassette (IMI229).
15
RV HR3-cbbO2-HR4
CCTAGAAATCTCGTCGATCTC
4th cloning fragment: PGK1p-cbbO2-ADH1t cassette (IMI229).
16
FW HR4-GroEL-HR5
ATCACTCTTACCAGGCTAGG
5th cloning fragment: TEF1p-groEL-ACT1t cassette (IMI229).
17
RV HR4-GroEL-HR5
CTGGACCTTAATCGTGTGCGCATCCTC
5th cloning fragment: TEF1p-groEL-ACT1t cassette (IMI229).
18
FW HR5-GroES-HR6
CCGTATAGCTTAATAGCCAGCTTTATC
6th cloning fragment: TPI1p-groES-PGI1t cassette (IMI229).
19
RV HR5-GroES-HR6
GCTATGACCATGATTACGCCAAGC
6th cloning fragment: TPI1p-groES-PGI1t cassette (IMI229).
20
FW HR6-LEU2-CAN1dwn
CCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCCTGTGAAGATCCCAGCAAAG
7th (IMI229) or 2nd (IMI232) cloning fragment: KlLEU2 cassette from pUG73.
21
RV LEU2 HR-CAN1
AGCTCATTGATCCCTTAAACTTTCTTTTCGGTGTATGACTTATGAGGGTGAGAATGCGAAATGGCGTGGAAATGTGATCAAAGGTAATAAAACGTCATATATCCGCAGGCTAACCGGAAC
7th (IMI229) or 2nd (IMI232) cloning fragment: KlLEU2 cassette from pUG73.
 
Primers used for verification of the in vivo assembled constructs
 
22
m-PCR-HR1-FW
GGCGATTAAGTTGGGTAACG
Diagnostic for assembly of plasmids pUDC075, pUDC099, and pUDC100, and integration in strain IMI229.
23
m-PCR-HR1-RV
AACTGAGCTCCAGCTGTACC
Diagnostic for assembly of plasmids pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
24
m-PCR-HR2-FW
ACGCGTGTACGCATGTAAC
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229
25
m-PCR-HR2-RV
GCGCGTGGCTTCCTATAATC
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229
26
m-PCR-HR3-FW
GTGAATGCTGGTCGCTATAC
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
27
m-PCR-HR3-RV
GTAAGCAGCAACACCTTCAG
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
28
m-PCR-HR4-FW
ACCTGACCTACAGGAAAGAG
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
29
m-PCR-HR4-RV
TGAAGTGGTACGGCGATGC
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
30
m-PCR-HR5-FW
ATAGCCACCCAAGGCATTTC
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
31
m-PCR-HR5-RV
CCGCACTTTCTCCATGAGG
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
32
m-PCR-HR6-FW
CGACGGTTACGGTGTTAAG
Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.
33 m-PCR-HR6-RV CTTCCGGCTCCTATGTTGTG Diagnostic for assembly of pUDC075, pUDC099, pUDC100, and integration in strain IMI229.