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Systematic tools for reprogramming plant gene expression in a simple model, Marchantia polymorpha.

Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M, Markel K, Delmans M, West A, Patron NJ, Haseloff J
ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
136067pCsAAcceptor plasmid for even level position 1, lacZ blue-white screening
136068pCsBAcceptor plasmid for even level position 2, lacZ blue-white screening
136069pCsCAcceptor plasmid for even level position 3, lacZ blue-white screening
136070pCsEAcceptor plasmid for even level position 4, lacZ blue-white screening
136071pCk1_spacerUse when making even level constructs (L2,...) if pCk1 position (position 1) has no construct assigned
136072pCk2_spacerUse when making even level constructs (L2,...) if pCk2 position (position 2) has no construct assigned
136073pCk3_spacerUse when making even level constructs (L2,...) if pCk3 position (position 3) has no construct assigned
136074pCk4_spacerUse when making even level constructs (L2,...) if pCk4 position (position 4) has no construct assigned
136075pCsA_spacerUse when making odd level constructs (L3,...) if pCsA position (position 1) has no construct assigned
136076pCsB_spacerUse when making odd level constructs (L3,...) if pCsB position (position 2) has no construct assigned
136077pCsC_spacerUse when making odd level constructs (L3,...) if pCsC position (position 3) has no construct assigned
136078pCsE_spacerUse when making odd level constructs (L3,...) if pCsE position (position 4) has no construct assigned
136079pUAP4Acceptor plasmid for L0 parts. Clone L0 parts into pUAP4 with SapI, lacZ blue-white screening. Release parts to assemble transcription units with BsaI.
136080CDS_nptIIConfers plant resistance to kanamycin
136081CDS_mALSConfers plant resistance to chlorsulfuron
136082CDS_hptIIConfers plant resistance to hygromycin
136083CDS_LUCFirefly luciferase
136084CDS_GUSFor histochemical _-glucuronidase activity assays. This GUS CDS does not have an intron.
136085CDS12_eGFPFP for N-term fusion with a CTAG
136086CDS12_eGFP-linkerFP for N-term fusion with a CTAG without linker
136087CDS12_mScarlet-IFP for N-term fusion with a CTAG
136088CDS12_mScarletI-linkerFP for N-term fusion with a CTAG without linker
136089CDS12_mTurquoise2FP for N-term fusion with a CTAG
136090CDS12_mTurquoise2-linkerFP for N-term fusion with a CTAG without linker
136091CDS12_mVenusFP for N-term fusion with a CTAG
136092CDS12_mVenus-linkerFP for N-term fusion with a CTAG without linker
136093CDS12_TP-MpSIG2Chloroplast transit peptide from MpSIG2. For N-term fusion with a CTAG
136094CDS12-MpER-TargCDS12 with ER targeting peptide to fuse with CTAG containing HDEL ER retention signal at the 3'end
136095CTAG_eGFPFP for C-term fusion with CDS12
136096CTAG_eGFP-GHtag-HDELFP with ER retention signal for C-term fusion with CDS12 with ER targeting signal peptide. Contains Gly-His tag
136097CTAG_eGFP-HDELFP with ER retention signal for C-term fusion with CDS12 with ER targeting signal peptide
136098CTAG_linker-eGFPFP for C-term fusion with CDS12 without linker
136099CTAG_linker-Lti6bPlasma Membrane targeting protein for C-term fusion with CDS12 without linker.
136100CTAG_linker-mTurquoise2FP for C-term fusion with CDS12 without linker
136101CTAG_linker-mVenusFP for C-term fusion with CDS12 without linker
136102CTAG_linker-N7Nuclear targeting peptide for C-term fusion with CDS12 without linker.
1361035UTR_MpNOP15UTR for cloning with PROM_MpNOP1
136104CTAG_mTurquoise2FP for C-term fusion with CDS12
136105CTAG_mTurquoise2-HDELFP with ER retention signal for C-term fusion with CDS12 with ER targeting signal peptide
136106CTAG_mVenusFP for C-term fusion with CDS12
136107CTAG_mVenus-HDELFP with ER retention signal for C-term fusion with CDS12 with ER targeting signal peptide
136108CTAG_NLS-SV40CTAG Nuclear localiation signal SV40 for fusion with CDS12
136109PROM_MpUBE2PROM for cloning with 5UTR_MpUBE2. PROM+5UTR_MpUBE2 has ubiquitous expression in early satges of gemmae development.
1361105UTR_MpUBE25UTR for cloning with PROM_MpUEB2. PROM+5UTR_MpUBE2 has ubiquitous expression in early stages of gemmae development.
136111PROM5_35SStrong promoter that drives expression everywhere in Mp gemmae but much weaker in the notches
136112PROM5_35Sx2Strong promoter that drives expression everywhere in Mp gemmae but much weaker in the notches
136113PROM5_MpEF1aStrong promoter from MpEF1a, that drives expression everywhere in Mp gemmae but stronger in the notches
1361143TERM_35S3TERM after CDS/ CDS12-CTAG
1361153TERM_Nos3TERM after CDS/ CDS12-CTAG
1361163TERM_Nos-35S3TERM after CDS/ CDS12-CTAG
136117PROM_MpNOP1PROM for cloning with 5UTR_MpNOP1
136118CTAG_mScarlet-IFP for C-term fusion with CDS12
1361193TERM_MpNOP13TERM after CDS/ CDS12-CTAG
136120PROM5_MpU6Mp U6 promoter (type III RNA polymerase promoter) to drive expression of gRNA for CRISPR/Cas9
136121CDS_Cas9-NLSPuchta Arabidopsis codon-optimised Cas9 for CRISPR
136122CDS12_Cas9-NLSPuchta Arabidopsis codon-optimised Cas9 for CRISPR. CDS12 for N-term fusion with a CTAG
136123L1_KanR-Ck1L1 in position 1 (next to LB in L2 construct), for KanR
136124L1_CsR-Ck1L1 in position 1 (next to LB in L2 construct), for CsR
136125L1_HyR-Ck1L1 in position 1 (next to LB in L2 construct), for HygR
136126L1_NOP1:mS-N7-Ck2L1 in position 2, example of weak promoter
136127L1_UBE2:mT-N7-Ck2L1 in position 2, control of expression
136128L1_UBE2:mT-N7-Ck3L1 in position 3, control of expression
136129L1_UBE2:eG-Lt-Ck3L1 in position 3, mark cell outline with eGFP
136130L1_UBE2:eG-Lt-Ck4L1 in position 4, mark cell outline with eGFP
136131L1_UBE2:mS-Lt-Ck3L1 in position 3, mark cell outline with mScarletI
136132L1_UBE2:mS-Lt-Ck4L1 in position 4, mark cell outline with mScarletI
136133L2_UBE2:mT-N7-CsAPlasmid to transform Mp with constitutive promoter driving mTurq to the nuclei. HygR
136134L2_UBE2:eG-Lt-CsAPlasmid to transform Mp with constitutive promoter marking cell outline with eGFP. HygR
136135L1_Cas9-Ck4L1 in position 4, for AtcoCas9 (Puchta) expression in L2 CRISPR vectors
136136L1_lacZgRNA-Ck3L1 in position 3, to clone gRNA target using BbsI, lacZ blue-white screening
136137L1_lacZgRNA-Ck2L1 in position 2, to clone gRNA target using BbsI, lacZ blue-white screening
136138L2_lacZgRNA-Cas9-CsAPlasmid to accept gRNA target with SapI, lacZ blue-white screening. Contains Cas9 and HygR.
136139L2_NOP1gRNA-Cas9-CsAPlasmid with NOP1-gRNA to tranform Mp as CRISPR control. Contains Cas9 and HygR
136140L2_2xNOP1gRNA-Cas9-CsAPlasmid with 2 different NOP1-gRNA. Targets 500 bp apart in the genome. To tranform Mp as CRISPR control. Contains Cas9 and HygR
136695pCk1Acceptor plasmid for odd level position 1, lacZ blue-white screening. Part of the OpenPlant toolkit.
136696pCk2Acceptor plasmid for odd level position 2, lacZ blue-white screening. Part of the OpenPlant toolkit.
136697pCk3Acceptor plasmid for odd level position 3, lacZ blue-white screening. Part of the OpenPlant toolkit.
136698pCk4Acceptor plasmid for odd level position 4, lacZ blue-white screening. Part of the OpenPlant toolkit.
1639313TERM_bba_b0015_rev_chloL0 part - terminator
1639323TERM_Ntpsba_rev_chloL0 part - terminator
163935CDS_aadA_chloL0 part - CDS
163936CDS_mTurqcp_chloL0 part - CDS
163937L1-Ck1-hs1-l-chloFor integration in the chloroplast trnG_trnfM region: HS1-L, genome position: 40216_ 41843, Genbank accession no. MH635409
163938L1-Ck1-hs2-l-chloFor integration in the chloroplast rbcL_trnR region: HS2-L, genome position: 56139_57662, Genbank accession no. MH635409
163939L1-Ck2-smR-chloL1 in position 2, for spectinomycin in chloroplast transformation
163940L1-Ck4-hs1-r-chloFor integration in the chloroplast trnG_trnfM region: HS1-R, genome position: 41847_ 43622, Genbank accession no. MH635409
163941L1-Ck4-hs2-r-chloFor integration in the chloroplast rbcL_trnR region: HS2-R, genome position: 57681- 59304, Genbank accession no. MH635409
163942pCk1chloAcceptor plasmid for odd level position 1
163943pCk2chloAcceptor plasmid for odd level position 2
163944pCk3chloAcceptor plasmid for odd level position 3
163945pCk4chloAcceptor plasmid for odd level position 4
163946pCsAchloAcceptor plasmid for even level position 1
163947pCsBchloAcceptor plasmid for even level position 2
163948pCsCchloAcceptor plasmid for even level position 3
163949pCsEchloAcceptor plasmid for even level position 4
163951PROM5_Mprnpo_chloL0 part - promoter
163952PROM5_Ntprrn_rev_chloL0 part - promoter
163953PROM5_Ntpsba_chloL0 part - promoter

Antibodies from Article