EPR Plasmid #871: pZmUBQ_zCas9i_P2A_GFP_4x_sg_ZMET
Sequence: download sequence
| Purpose of plasmid | Plasmid harbouring, under ZmUbiquitin promoter, Cas9 with introns fused to eGFP with a P2A signal and the cassettes to express 4 specific sgRNAs for spZmet.It can be used for Crispr editing of SpZMET. |
| Depositing Lab |
Arturo Mari-Ordonez Gregor Mendel Institute of Molecular Plant Biology |
| Depositing Scientist | Veronica Barragan-Borrero |
| Antibiotic Resistance | Spectinomycin |
| Bacterial Growth Temp | 37 |
| Copy Number | High |
| Selectable Marker | Hygromycin |
| Final expression system: | Plants, monocots |
| Vector type: | T-DNA binary vector |
| Insert type: | cDNA+sgRNA |
| Insert gene: | Cas9 and 4 cassettes RNA polIII promoter_sgRNA Zmet_sgRNAscaffold |
| Reference |
Strain, procedures, and tools for reproducible genetic transformation and genome editing of Spirodela polyrhiza (L.) Schleid. Barragan-Borrero, V.; de Santana Lopes, A.; Rodrigues-Batista, E. D.; Hofer, M.; Elias, R.; Chakrab... DOI |
| MTA and Terms |
MTA (PDF download) |
How to cite this reagent:
In the Materials & Methods:
pZmUBQ_zCas9i_P2A_GFP_4x_sg_ZMET was a gift from Arturo Mari-Ordonez (European Plasmid Repository plasmid #871).
In the References:
Barragan-Borrero, V.; de Santana Lopes, A.; Rodrigues-Batista, E. D.; Hofer, M.; Elias, R.; Chakrab.... Strain, procedures, and tools for reproducible genetic transformation and genome editing of Spirodela polyrhiza (L.) Schleid. . doi: 10.1101/2025.08.28.672806
pZmUBQ_zCas9i_P2A_GFP_4x_sg_ZMET was a gift from Arturo Mari-Ordonez (European Plasmid Repository plasmid #871).
In the References:
Barragan-Borrero, V.; de Santana Lopes, A.; Rodrigues-Batista, E. D.; Hofer, M.; Elias, R.; Chakrab.... Strain, procedures, and tools for reproducible genetic transformation and genome editing of Spirodela polyrhiza (L.) Schleid. . doi: 10.1101/2025.08.28.672806
Sequence: download sequence
CTCTTTTCTCTTAGGTTTACCCGCCAATATATCCTGTCAAACACTGATAGTTTCTGCAGTGCAGCGTGACCCGGTCGTGCCCCTCTCTAGAGATAATGAGCATTGCATGTCTAAGTTATAAAAAATTACCACATATTTTTTTTGTCACACTTGTTTGAAGTGCAGTTTATCTATCTTTATACATATATTTAAACTTTACTCTACGAATAATATAATCTATAGTACTACAATAATATCAGTGTTTTAGAGAATCATATAAATGAACAGTTAGACATGGTCTAAAGGACAATTGAGTATTTTGACAACAGGACTCTACAGTTTTATCTTTTTAGTGTGCATGTGTTCTCCTTTTTTTTTGCAAATAGCTTCACCTATATAATACTTCATCCATTTTATTAGTACATCCATTTAGGGTTTAGGGTTAATGGTTTTTATAGACTAATTTTTTTAGTACATCTATTTTATTCTATTTTAGCCTCTAAATTAAGAAAACTAAAACTCTATTTTAGTTTTTTTATTTAATAATTTAGATATAAAATAGAATAAAATAAAGTGACTAAAAATTAAACAAATACCCTTTAAGAAATTAAAAAAACTAAGGAAACATTTTTCTTGTTTCGAGTAGATAATGCCAGCCTGTTAAACGCCGTCGACGAGTCTAACGGACACCAACCAGCGAACCAGCAGCGTCGCGTCGGGCCAAGCGAAGCAGACGGCACGGCATCTCTGTCGCTGCCTCTGGACCCCTCTCGAGAGTTCCGCTCCACCGTTGGACTTGCGTGGCGGAGCGGCAGACGTGAGCCGGCACGGCAGGCGGCCTCCTCCTCCTCTCACGGCACCGGCAGCTACGGGGGATTCCTTTCCCACCGCTCCTTCGCTTTCCCTTCCTCGCCCGCCGTAATAAATAGACACCCCCTCCACACCCTCTTTCCCCAACCTCGTGTTGTTCGGAGCGCACACACACACAACCAGATCTCCCCCAAATCCACCCGTCGGCACCTCCGCTTCAAGGTACGCCGCTCGTCCTCCCCCCCCCCTCTCTACCTTCTCTAGATCGGCGTTCCGGTCCATGGTTAGGGCCCGGTAGTTCTACTTCTGTTCATGTTTGTGTTAGATCCGTGTTTGTGTTAGATCCGTGCTGCTAGCGTTCGTACACGGATGCGACCTGTACGTCAGACACGTTCTGATTGCTAACTTGCCAGTGTTTCTCTTTGGGGAATCCTGGGATGGCTCTAGCCGTTCCGCAGACGGGATCGATTTCATGATTTTTTTTGTTTCGTTGCATAGGGTTTGGTTTGCCCTTTTCCTTTATTTCAATATATGCCGTGCACTTGTTTGTCGGGTCATCTTTTCATGCTTTTTTTTGTCTTGGTTGTGATGATGTGGTCTGGTTGGGCGGTCGTTCTAGATCGGAGTAGAATTCTGTTTCAAACTACCTGGTGGATTTATTAATTTTGGATCTGTATGTGTGTGCCATACATATTCATAGTTACGAATTGAAGATGATGGATGGAAATATCGATCTAGGATAGGTATACATGTTGATGCGGGTTTTACTGATGCATATACAGAGATGCTTTTTGTTCGCTTGGTTGTGATGATGTGGTGTGGTTGGGCGGTCGTTCATTCGTTCTAGATCGGAGTAGAATACTGTTTCAAACTACCTGGTGTATTTATTAATTTTGGAACTGTATGTGTGTGTCATACATCTTCATAGTTACGAGTTTAAGATGGATGGAAATATCGATCTAGGATAGGTATACATGTTGATGTGGGTTTTACTGATGCATATACATGATGGCATATGCAGCATCTATTCATATGCTCTAACCTTGAGTACCTATCTATTATAATAAACAAGTATGTTTTATAATTATTTTGATCTTGATATACTTGGATGATGGCATATGCAGCAGCTATATGTGGATTTTTTTAGCCCTGCCTTCATACGCTATTTATTTGCTTGGTACTGTTTCTTTTGTCGATGCTCACCCTGTTGTTTGGTGTTACTTCTGCAGAAACTAGtGCTCGAtcCCATGGCTCCTGCTGCCAAAAGAGTTAAACTCGACTCAGCTGCCGCAATGGACAAGAAGTACAGCATTGGACTTGATATTGGTACGAACTCAGTTGGGTGGGCCGTTATCACCGATGAATACAAGGTACCTTCGAAGAAATTTAAAGTGCTGGGCAACACAGATAGGCACAGCATTAAGAAGAACTTGATCGGAGCTCTGCTCTTTGACTCTGGAGAAACCGCGGAGGCGACAAGGCTTAAACGTACTGCGAGGAGAAGGTACACTCGCAGGAAGAACAGAATCTGTTATCTCCAAGAGATCTTTAGCAACGAGATGGCGAAGGTAAGGATTTTTATGATATAGTATGCTTATGTATTTTGTACTGAAAGCATATCCTGCTTCATTGGGATATTACTGAAAGCATTTAACTACATGTAAACTCACTTGATGATCAATAAACTTGATTTTGCAGGTTGACGACTCGTTCTTCCATCGCCTCGAGGAATCCTTCCTGGTAGAGGAAGATAAGAAACACGAGCGTCACCCCATCTTTGGGAATATTGTTGACGAAGTAGCCTATCATGAAAAGTATCCGACTATATACCACCTTCGCAAGAAGCTGGTGGACTCAACCGATAAGGCAGACCTTCGGCTCATATACCTGGCTCTCGCGCACATGATAAAGTTTCGTGGCCATTTCTTGATCGAAGGGGACCTCAACCCGGATAACTCCGATGTGGATAAACTGTTCATTCAGCTCGTCCAAACCTACAATCAGCTGTTCGAGGAGAACCCCATCAATGCATCAGGTAACATTCCTTAGTTACCTTTCTTTTCTTTTTCCATCATAAGTTTATAGATTGTACATGCTTTGAGATTTTTCTTTGCAAACAATCTCAGGTGTCGACGCCAAGGCAATACTGTCTGCCAGACTTTCGAAGTCCAGACGGCTTGAGAATCTGATCGCTCAATTGCCAGGCGAGAAGAAGAACGGCTTGTTCGGGAATCTGATTGCACTGTCTCTGGGCCTCACCCCTAACTTCAAAAGCAACTTTGACCTCGCCGAGGACGCGAAGCTGCAGCTGTCAAAGGATACATACGATGATGATCTGGACAATCTGCTCGCCCAAATAGGTGCTCTTGAAATTGGAACTCTTCTTTTGTTGTCTAAACCTATCAATTTCTTTGCGGAAATTTATTTGAAGCTGTAGAGTTAAAATTGAGTCTTTTAAACTTTTGTAGGTGATCAGTATGCCGACCTGTTCTTGGCTGCCAAGAATCTGTCAGACGCTATCTTGCTCAGTGACATTCTGCGGGTCAACACGGAGATAACCAAAGCGCCACTTAGCGCCTCCATGATCAAGAGGTACGACGAGCATCACCAGGATCTGACCCTTCTGAAGGCTTTGGTTCGCCAGCAACTCCCCGAGAAGTACAAGGAGATTTTCTTTGACCAATCGAAGAATGGCTACGCAGGGTACATTGATGGAGGTAAGTTGTTACTTATGATTGTTTTCCTCTCTGCTACATGTATTTTGTTGTTCATTTCTGTAAGATATAAGAATTGAGTTTTCCTCTGATGATATTATTAGGTGCAAGTCAGGAGGAATTCTACAAATTCATCAAGCCTATTCTGGAAAAGATGGACGGTACAGAGGAGCTGCTCGTTAAATTGAACCGCGAAGATTTGCTTCGGAAGCAGCGTACCTTCGACAATGGCAGCATACCGCACCAGATCCACCTCGGTGAGCTGCATGCTATCTTGAGGAGGCAAGAGGACTTCTATCCGTTCCTGAAAGACAACAGAGAGAAGATTGAAAAGATCCTCACGTTCCGCATTCCCTACTATGTAGGTTAGTATCATATGAAGAAATACCTAGTTTCAGTTGATGAATGCTATTTTCTGACCTCAGTTGTTCTCTTTTGAGAATTATTTCTTTTCTAATTTGCCTGATTTTTCTATTAATTCATTAGGTCCACTCGCACGCGGGAACTCGCGGTTTGCGTGGATGACACGCAAATCCGAGGAGACTATCACGCCTTGGAACTTCGAAGAGGTCGTGGACAAGGGTGCGAGTGCACAGTCCTTCATCGAAAGGATGACCAACTTCGATAAGAATCTCCCAAATGAGAAAGTCCTGCCCAAGCATAGTCTCCTGTACGAATACTTCACGGTCTACAACGAGCTGACGAAGGTGAAATATGTGACGGAGGGGATGCGCAAACCGGCCTTCCTGTCAGGTAAATCCTGGTCCACACTTTTACGATAAAAACACAAGATTTTAAACTATGAACTGATCAATAATCATTCCTAAAAGACCACACTTTTGTTTTGTTTCTAAAGTAATTTTTACTGTTATAACAGGTGAGCAGAAGAAGGCCATTGTCGATCTCTTGTTCAAAACCAATCGGAAGGTCACTGTGAAACAGCTTAAAGAGGACTACTTTAAGAAGATCGAATGCTTTGATTCTGTGGAAATCAGCGGCGTTGAGGATAGGTTCAATGCCTCTCTTGGCACATACCATGACCTGTTGAAAATCATCAAGGACAAGGACTTCCTTGACAACGAGGAGAACGAGGACATCCTCGAGGACATCGTGCTGACTCTCACGCTGTTTGAGGACAGAGAAATGATCGAGGAGCGCCTTAAGACTTATGCGCATCTGTTCGATGACAAGGTCATGAAGCAGTTGAAGAGGAGGAGATATACAGGTAAGAGGTCAAAAGGTTTCCGCAATGATCCCTCTTTTTTTGTTTCTCTAGTTTCAAGAATTTGGGTATATGACTAACTTCTGAGTGTTCCTTGATGCATATTTGTGATGAGACAAATGTTTGTTCTATGTTTTAGGTTGGGGAAGGCTCTCCAGGAAGCTCATCAACGGCATCCGCGACAAGCAATCCGGCAAGACTATACTGGACTTTCTCAAATCCGACGGTTTTGCGAATCGGAACTTCATGCAGCTTATTCACGATGACTCACTGACCTTCAAAGAAGATATCCAGAAGGCCCAAGTGTCAGGTCAGGGCGATAGCCTTCACGAACACATAGCCAACCTGGCTGGATCGCCAGCTATAAAGAAGGGCATACTGCAGACAGTGAAGGTTGTGGATGAGCTGGTGAAGGTAAGTTCTGCATTTGGTTATGCTCCTTGCATTTTAGGTGTTCGTCGCACTTCCATTTCCATGAATAGCTAAGATTTTTTTTCTCTGCATTCATTCTTCTTGCCTCAGTTCTAACTGTTTGTGGTATTTTTGTTTTAATTATTGCTACAGGTCATGGGCCGCCATAAGCCGGAGAACATCGTCATCGAGATGGCGAGGGAAAACCAGACGACTCAGAAAGGGCAGAAGAACTCACGGGAGCGCATGAAGCGGATAGAGGAAGGCATCAAGGAGCTTGGGAGTCAGATTCTGAAAGAGCACCCAGTCGAAAATACTCAACTCCAGAACGAGAAGCTGTACCTCTATTACCTCCAGAATGGGAGAGATATGTACGTCGACCAAGAGCTCGACATTAACAGACTCTCCGACTATGATGTGGATCACATTGTCCCTCAATCTTTCCTGAAGGACGATAGTATTGACAACAAGGTAAAGCAACTGTGTTTTAATCAATTTCTTGTCAGGATATATGGATTATAACTTAATTTTTGAGAAATCTGTAGTATTTGGCGTGAAATGAGTTTGCTTTTTGGTTTCTCCCGTGTTATAGGTCCTTACGCGCTCAGACAAGAACCGCGGAAAATCCGACAATGTACCCAGCGAGGAGGTTGTGAAGAAGATGAAGAACTATTGGAGGCAGCTTTTGAATGCTAAGCTCATAACCCAACGGAAATTCGACAATCTCACGAAGGCAGAAAGGGGCGGACTGTCTGAGCTCGACAAAGCCGGCTTCATCAAGCGCCAGTTGGTTGAAACTCGTCAGATTACGAAACATGTGGCCCAGATACTCGATTCGCGTATGAATACGAAGTATGATGAGAATGACAAACTTATCAGGGAGGTAAAGGTAAAGTTTCCAACTTTCCTTTACCATATCAAACTAAAGTTCGAAACTTTTTATTTGATCAACTTCAAGGCCACCCGATCTTTCTATTCCTGATTAATTTGTGATGAATCCATATTGACTTTTGATGGTTACGCAGGTGATCACCCTCAAGAGCAAACTGGTTAGTGACTTCCGGAAGGACTTCCAGTTTTACAAGGTTCGCGAGATCAACAACTACCATCATGCCCATGACGCCTACCTGAACGCCGTTGTTGGCACTGCTCTCATCAAGAAGTATCCGAAACTGGAGTCTGAGTTTGTGTACGGGGATTACAAGGTGTACGACGTTAGGAAGATGATCGCGAAGTCAGAACAAGAGATCGGCAAGGCTACCGCGAAATACTTCTTTTACTCGAATATCATGAACTTCTTCAAGACAGAGATCACTCTGGCGAATGGTGAAATCCGGAAGAGGCCTCTGATCGAGACAAATGGCGAAACAGGTCTGTCTTTCCTATTTCATATGTTTAATCCTAGGAATTTGATCAATTGATTGTATGTATGTCGATCCCAAGACTTTCTTGTTCACTTATATCTTAACTCTCTCTTTGCTGTTTCTTGCAGGTGAGATTGTCTGGGATAAGGGCAGGGATTTTGCGACTGTGCGTAAGGTTCTCAGCATGCCCCAAGTCAACATAGTCAAGAAAACGGAGGTTCAAACCGGTGGTTTCTCCAAGGAGTCCATTCTCCCTAAGCGCAACTCCGACAAACTGATTGCGAGGAAGAAGGATTGGGATCCGAAGAAATACGGAGGCTTTGATAGCCCTACCGTGGCATACAGCGTACTGGTAGTGGCCAAGGTGGAGAAGGGCAAGAGCAAGAAACTGAAAAGCGTCAAGGAACTGCTTGGAATTACCATAATGGAAAGGTCCTCGTTCGAGAAGAATCCGATCGACTTCCTCGAGGCTAAAGGTAAAATATTGGATGCCAGACGATATTCTTTCTTTTGATTTGTAACTTTTTCCTGTCAAGGTCGATAAATTTTATTTTTTTTGGTAAAAGGTCGATAATTTTTTTTTGGAGCCATTATGTAATTTTCCTAATTAACTGAACCAAAATTATACAAACCAGGTTACAAAGAGGTGAAGAAAGACCTCATTATCAAACTGCCCAAGTATTCGCTTTTCGAATTGGAAAATGGCAGAAAACGCATGCTGGCATCTGCCGGAGAACTGCAGAAGGGCAACGAGCTGGCATTGCCCAGTAAGTACGTCAACTTCCTGTACTTGGCCTCACACTATGAGAAGCTGAAGGGGTCACCAGAGGACAACGAGCAGAAGCAGTTGTTTGTCGAGCAGCACAAGCACTATCTTGATGAGATCATAGAGCAGATCAGCGAATTTTCCAAGCGGGTCATTCTTGCAGACGCTAACCTCGATAAGGTAAGGACTTCTCATGAATATTAGTGGCAGATTAGTGTTGTTAAAGTCTTTGGTTAGATAATCGATGCCTCCTAATTGTCCATGTTTTACTGGTTTTCTACAATTAAAGGTGCTTTCCGCGTACAACAAGCACAGAGATAAGCCGATAAGGGAACAAGCGGAAAACATCATCCACCTGTTCACACTGACCAATCTGGGAGCCCCAGCAGCCTTTAAGTACTTCGATACCACTATCGACAGAAAGCGCTACACATCAACCAAGGAAGTGTTGGACGCTACCCTTATTCACCAATCTATTACAGGGCTCTATGAGACAAGGATAGATCTGTCGCAGTTGGGTGGTGACTCTAGGGCTGACCCAAAGAAGAAGCGTAAAGTCGGATCCGGAGCAACCAACTTTAGCCTGCTCAAGCAAGCAGGAGATGTTGAGGAAAATCCTGGCCCCaTGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCTTCACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCACGGCATGGACGAGCTGTACAAGtaagcaCGGTAaGATATCCCGCGGCCATGGCGGCCGGGAGCATGCGACGTCGATCTAACTGACTAGCCGCGGCCATGCTAGAGTCCGCAAAAATCACCAGTCTCTCTCTACAAATCTATCTCTCTCTATTTTTCTCCAGAATAATGTGTGAGTAGTTCCCAGATAAGGGAATTAGGGTTCTTATAGGGTTTCGCTCATGTGTTGAGCATATAAGAAACCCTTAGTATGTATTTGTATTTGTAAAATACTTCTATCAATAAAATTTCTAATcCCTAAAACCAAAATCCAGTGACCTGCAaAGCTTGAAACTGAAGGCGGGAAACGACAATCTGATCATGAGCGGAGAATTAAGGGAGTCACGTTATGACCCCCGCCGATGACGCGGGACAAGCCGTTTTACGTTTGGAACTGACAGAACCGCAACGTTGAAGGAGCCACTCAGCCGCGGGTTTCTGGAGTTTAATGAGCTAAGCACATACGTCAGAAACCATTATTGCGCGTTCAAAAGTCGCCTAAGGTCACTATCAGCTAGCAAATATTTCTTGTCAAAAATGCTCCACTGACGTTCCATAAATTCCCCTCGGTATCCAATTAGAGTCTCATATTCACTCTCAATCCAAATAATCTGCACCGGATCCGCTAGAGGATCTCGACCTGCAAGATCCCGGGGGGCAATGAGATATGAAAAAGCCTGAACTCACCGCGACGTCTGTCGAGAAGTTTCTGATCGAAAAGTTCGACAGCGTCTCCGACCTGATGCAGCTCTCGGAGGGCGAAGAATCTCGTGCTTTCAGCTTCGATATAGGAGGGCGTGGATATGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCCGCGCTCCCGATTCCGGAAGTGCTTGACATTGGGGAATTCAGCGAGAGCCTGACCTATTGCATCTCCCGCCGTGCACAGGGTGTCACGTTGCAAGACCTGCCTGAAACCGAACTGCCCGCTGTTCTGCAGCCGGTCGCGGAGGCCATGGATGCGATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTTCGGCCCATTCGGACCGCAAGGAATCGGTCAATACACTACATGGCGTGATTTCATATGCGCGATTGCTGATCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCGTCAGTGCGTCCGTCGCGCAGGCTCTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGAAGTCCGGCACCTCGTGCACGCGGATTTCGGCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTGACTGGAGCGAGGCGATGTTCGGGGATTCCCAATACGAGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGTATGGAGCAGCAGACGCGCTACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCCGGGCGTATATGCTCCGCATTGGTCTTGACCAACTCTATCAGAGCTTGGTTGACGGCAATTTCGATGATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATCGTCCGATCCGGAGCCGGGACTGTCGGGCGTACACAAATCGCCCGCAGAAGCGCGGCCGTCTGGACCGATGGCTGTGTAGAAGTACTCGCCGATAGTGGAAACCGACGCCCCAGCACTCGTCCGGGATCTTGGAGGTGATGTAACATGATCACAAGCTGATCCCCCGAATTTCCCCGATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATCGGGAATTGATCCCCCCTCGACAGCTTGCATGCCAGCTTGGGCTGCAGGTCGACGCGtGTTACTAGATCGGGAGCACCGGTAAGGCGCGCCGTAGTGCTCGtAcTGGAATCGGCAGCAAAGGAAGGAATCTTTAAACATACGAACAGATCACTTAAAGTTCTTCTGAAGCAACTTAAAGTTATCAGGCATGCATGGATCTTGGAGGAATCAGATGTGCAGTCAGGGACCATAGCACAAGACAGGCGTCTTCTACTGGTGCTACCAGCAAATGCTGGAAGCCGGGAACACTGGGTACGTTGGAAACCACGTGTGATGTGAAGGAGTAAGATAAACTGTAGGAGAAAAGCATTTCGTAGTGGGCCATGAAGCCTTTCAGGACATGTATTGCAGTATGGGCCGGCCCATTACGCAATTGGACGACAACAAAGGCTAGTATTAGTACCACCTCGGCTATCCACATAGATCAAAGCTGGTTTAAAAGAGTTGTGCAGATGATCCGTGGCAGGCGAAATGCCACTACCGCGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTTCAAGAGCTTGGAGTGGATGGACCCTGACACTGGAATCGGCAGCAAAGGACGCGTTGACATTGTAGGACTATATTGCTCTAATAAAGGAGGCAGCTATGCTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCTAATTTTTTCCTGTAGTTTTCCCACAACCATTTTTTACCATCCGAATGATAGGATAGGAAAAATATCCAAGTGAACAGTATTCCTATAAAATTCCCGTAAAAAGCCTGCAATCCGAATGAGCCCTGAAGTCTGAACTAGCCGGTCACCTGTACAGGCTATCGAGATGCCATACAAGAGACGGTAGTAGGAACTAGGAAGACGATGGTTGATTCGTCAGGCGAAATCGTCGTCCTGCAGTCGCATCTATGGGCCTGGACGGAATAGGGGAAAAAGTTGGCCGGATAGGAGGGAAAGGCCCAGGTGCTTACGTGCGAGGTAGGCCTGGGCTCTCAGCACTTCGATTCGTTGGCACCGGGGTAGGATGCAATAGAGAGCAACGTTTAGTACCACCTCGCTTAGCTAGAGCAAACTGGACTGCCTTATATGCGCGGGTGCTGGCTTGGCTGCCGTCGTTCGGCGTCGCAGAAGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTTCAAGAGCTTGGAGTGGATGGAGTGAAGACTTTGGAATCGGCAGCAAAGGATGCAAGAACGAACTAAGCCGGACAAAAAAAAAAGGAGCACATATACAAACCGGTTTTATTCATGAATGGTCACGATGGATGATGGGGCTCAGACTTGAGCTACGAGGCCGCAGGCGAGAGAAGCCTAGTGTGCTCTCTGCTTGTTTGGGCCGTAACGGAGGATACGGCCCACGAGCGTGTACTACCGCGCGGGATGCCGCTGGGCGCTGCGGGGGCCGTTGGATGGGGATCGGTGGGTCGCGGGAGCGTTGAGGGGAGACAGGTTTAGTACCACCTCGCCTACCGAACAATGAAGAACCCACCTTATAACCCCGCGCGCTGCCGCTTGTGTTGCAAGGGTCCACCCTTCCGTTGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTTCAAGAGCTTGGAGTGGATGGAAAGGACTACATGGAATCGGCAGCAAAGGACTCATTAGCGGTATGCATGTTGGTAGAAGTCGGAGATGTAAATAATTTTCATTATATAAAAAAGGTACTTCGAGAAAAATAAATGCATACGAATTAATTCTTTTTATGTTTTTTAAACCAAGTATATAGAATTTATTGATGGTTAAAATTTCAAAAATATGACGAGAGAAAGGTTAAACGTACGGCATATACTTCTGAACAGAGAGGGAATATGGGGTTTTTGTTGCTCCCAACAATTCTTAAGCACGTAAAGGAAAAAAGCACATTATCCACATTGTACTTCCAGAGATATGTACAGCATTACGTAGGTACGTTTTCTTTTTCTTCCCGGAGAGATGATACAATAATCATGTAAACCCAGAATTTAAAAAATATTCTTTACTATAAAAATTTTAATTAGGGAACGTATTATTTTTTACATGACACCTTTTGAGAAAGAGGGACTTGTAATATGGGACAAATGAACAATTTCTAAGAAATGGGCATATGACTCTCAGTACAATGGACCAAATTCCCTCCAGTCGGCCCAGCAATACAAAGGGAAAGAAATGAGGGGGCCCACAGGCCACGGCCCACTTTTCTCCGTGGTGGGGAGATCCAGCTAGAGGTCCGGCCCACAAGTGGCCCTTGCCCCGTGGGACGGTGGGATTGCAGAGCGCGTGGGCGGAAACAACAGTTTAGTACCACCTCGCTCACGCAACGACGCGACCACTTGCTTATAAGCTGCTGCGCTGAGGCTCAGATCTTCCTTCCGCTTCCTCGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTTCAAGAGCTTGGAGTGGATGGAgTaCGGTAaAGCTTgatgctcgagGGCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTaCGCGTCAATTGATATCGATCCAGGCCTCCCAGCTTTCGTCCGTATCATCGGTTTCGACAACGTTCGTCAAGTTCAATGCATCAGTTTCATTGCCCACACACCAGAATCCTACTAAGTTTGAGTATTATGGCATTGGAAAAGCTGTTTTCTTCTATCATTTGTTCTGCTTGTAATTTACTGTGTTCTTTCAGTTTTTGTTTTCGGACATCAAAATGCAAATGGATGGATAAGAGTTAATAAATGATATGGTCCTTTTGTTCATTCTCAAATTATTATTATCTGTTGTTTTTACTTTAATGGGTTGAATTTAAGTAAGAAAGGAACTAACAGTGTGATATTAAGGTGCAATGTTAGACATATAAAACAGTCTTTCACCTCTCTTTGGTTATGTCTTGAATTGGTTTGTTTCTTCACTTATCTGTGTAATCAAGTTTACTATGAGTCTATGATCAAGTAATTATGCAATCAAGTTAAGTACAGTATAGGCTTTTTGTGTCGAGGGGGTACCGAGTCGAGGAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGGGTACCGAGCTCGAATTCAATTCGGCGTTAATTCAGTACATTAAAAACGTCCGCAATGTGTTATTAAGTTGTCTAAGCGTCAATTTGTTTACACCACAATATATCCTGCCACCAGCCAGCCAACAGCTCCCCGACCGGCAGCTCGGCACAAAATCACCACTCGATACAGGCAGCCCATCAGTCCGGGACGGCGTCAGCGGGAGAGCCGTTGTAAGGCGGCAGACTTTGCTCATGTTACCGATGCTATTCGGAAGAACGGCAACTAAGCTGCCGGGTTTGAAACACGGATGATCTCGCGGAGGGTAGCATGTTGATTGTAACGATGACAGAGCGTTGCTGCCTGTGATCAATTCGGGCACGAACCCAGTGGACATAAGCCTGTTCGGTTCGTAAGCTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGGGGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAAACATCATGGGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAATGTCTAGCTAGAAATTCGTTCAAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAAGCGTTAGATGCACTAAGCACATAATTGCTCACAGCCAAACTATCAGGTCAAGTCTGCTTTTATTATTTTTAAGCGTGCATAATAAGCCCTACACAAATTGGGAGATATATCATGCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGGCAGGGTGCCTTGATGTGGGCGCCGGCGGTCGAGTGGCGACGGCGCGGCTTGTCCGCGCCCTGGTAGATTGCCTGGCCGTAGGCCAGCCATTTTTGAGCGGCCAGCGGCCGCGATAGGCCGACGCGAAGCGGCGGGGCGTAGGGAGCGCAGCGACCGAAGGGTAGGCGCTTTTTGCAGCTCTTCGGCTGTGCGCTGGCCAGACAGTTATGCACAGGCCAGGCGGGTTTTAAGAGTTTTAATAAGTTTTAAAGAGTTTTAGGCGGAAAAATCGCCTTTTTTCTCTTTTATATCAGTCACTTACATGTGTGACCGGTTCCCAATGTACGGCTTTGGGTTCCCAATGTACGGGTTCCGGTTCCCAATGTACGGCTTTGGGTTCCCAATGTACGTGCTATCCACAGGAAAagaccTTTTCGACCTTTTTCCCCTGCTAGGGCAATTTGCCCTAGCATCTGCTCCGTACATTAGGAACCGGCGGATGCTTCGCCCTCGATCAGGTTGCGGTAGCGCATGACTAGGATCGGGCCAGCCTGCCCCGCCTCCTCCTTCAAATCGTACTCCGGCAGGTCATTTGACCCGATCAGCTTGCGCACGGTGAAACAGAACTTCTTGAACTCTCCGGCGCTGCCACTGCGTTCGTAGATCGTCTTGAACAACCATCTGGCTTCTGCCTTGCCTGCGGCGCGGCGTGCCAGGCGGTAGAGAAAACGGCCGATGCCGGGATCGATCAAAAAGTAATCGGGGTGAACCGTCAGCACGTCCGGGTTCTTGCCTTCTGTGATCTCGCGGTACATCCAATCAGCTAGCTCGATCTCGATGTACTCCGGCCGCCCGGTTTCGCTCTTTACGATCTTGTAGCGGCTAATCAAGGCTTCACCCTCGGATACCGTCACCAGGCGGCCGTTCTTGGCCTTCTTCGTACGCTGCATGGCAACGTGCGTGGTGTTTAACCGAATGCAGGTTTCTACCAGGTCGTCTTTCTGCTTTCCGCCATCGGCTCGCCGGCAGAACTTGAGTACGTCCGCAACGTGTGGACGGAACACGCGGCCGGGCTTGTCTCCCTTCCCTTCCCGGTATCGGTTCATGGATTCGGTTAGATGGGAAACCGCCATCAGTACCAGGTCGTAATCCCACACACTGGCCATGCCGGCCGGCCCTGCGGAAACCTCTACGTGCCCGTCTGGAAGCTCGTAGCGGATCACCTCGCCAGCTCGTCGGTCACGCTTCGACAGACGGAAAACGGCCACGTCCATGATGCTGCGACTATCGCGGGTGCCCACGTCATAGAGCATCGGAACGAAAAAATCTGGTTGCTCGTCGCCCTTGGGCGGCTTCCTAATCGACGGCGCACCGGCTGCCGGCGGTTGCCGGGATTCTTTGCGGATTCGATCAGCGGCCGCTTGCCACGATTCACCGGGGCGTGCTTCTGCCTCGATGCGTTGCCGCTGGGCGGCCTGCGCGGCCTTCAACTTCTCCACCAGGTCATCACCCAGCGCCGCGCCGATTTGTACCGGGCCGGATGGTTTGCGACCGCTCACGCCGATTCCTCGGGCTTGGGGGTTCCAGTGCCATTGCAGGGCCGGCAGACAACCCAGCCGCTTACGCCTGGCCAACCGCCCGTTCCTCCACACATGGGGCATTCCACGGCGTCGGTGCCTGGTTGTTCTTGATTTTCCATGCCGCCTCCTTTAGCCGCTAAAATTCATCTACTCATTTATTCATTTGCTCATTTACTCTGGTAGCTGCGCGATGTATTCAGATAGCAGCTCGGTAATGGTCTTGCCTTGGCGTACCGCGTACATCTTCAGCTTGGTGTGATCCTCCGCCGGCAACTGAAAGTTGACCCGCTTCATGGCTGGCGTGTCTGCCAGGCTGGCCAACGTTGCAGCCTTGCTGCTGCGTGCGCTCGGACGGCCGGCACTTAGCGTGTTTGTGCTTTTGCTCATTTTCTCTTTACCTCATTAACTCAAATGAGTTTTGATTTAATTTCAGCGGCCAGCGCCTGGACCTCGCGGGCAGCGTCGCCCTCGGGTTCTGATTCAAGAACGGTTGTGCCGGCGGCGGCAGTGCCTGGGTAGCTCACGCGCTGCGTGATACGGGACTCAAGAATGGGCAGCTCGTACCCGGCCAGCGCCTCGGCAACCTCACCGCCGATGCGCGTGCCTTTGATCGCCCGCGACACGACAAAGGCCGCTTGTAGCCTTCCATCCGTGACCTCAATGCGCTGCTTAACCAGCTCCACCAGGTCGGCGGTGGCCCATATGTCGTAAGGGCTTGGCTGCACCGGAATCAGCACGAAGTCGGCTGCCTTGATCGCGGACACAGCCAAGTCCGCCGCCTGGGGCGCTCCGTCGATCACTACGAAGTCGCGCCGGCCGATGGCCTTCACGTCGCGGTCAATCGTCGGGCGGTCGATGCCGACAACGGTTAGCGGTTGATCTTCCCGCACGGCCGCCCAATCGCGGGCACTGCCCTGGGGATCGGAATCGACTAACAGAACATCGGCCCCGGCGAGTTGCAGGGCGCGGGCTAGATGGGTTGCGATGGTCGTCTTGCCTGACCCGCCTTTCTGGTTAAGTACAGCGATAACCTTCATGCGTTCCCCTTGCGTATTTGTTTATTTACTCATCGCATCATATACGCAGCGACCGCATGACGCAAGCTGTTTTACTCAAATACACATCACCTTTTTAGACGGCGGCGCTCGGTTTCTTCAGCGGCCAAGCTGGCCGGCCAGGCCGCCAGCTTGGCATCAGACAAACCGGCCAGGATTTCATGCAGCCGCACGGTTGAGACGTGCGCGGGCGGCTCGAACACGTACCCGGCCGCGATCATCTCCGCCTCGATCTCTTCGGTAATGAAAAACGGTTCGTCCTGGCCGTCCTGGTGCGGTTTCATGCTTGTTCCTCTTGGCGTTCATTCTCGGCGGCCGCCAGGGCGTCGGCCTCGGTCAATGCGTCCTCACGGAAGGCACCGCGCCGCCTGGCCTCGGTGGGCGTCACTTCCTCGCTGCGCTCAAGTGCGCGGTACAGGGTCGAGCGATGCACGCCAAGCAGTGCAGCCGCCTCTTTCACGGTGCGGCCTTCCTGGTCGATCAGCTCGCGGGCGTGCGCGATCTGTGCCGGGGTGAGGGTAGGGCGGGGGCCAAACTTCACGCCTCGGGCCTTGGCGGCCTCGCGCCCGCTCCGGGTGCGGTCGATGATTAGGGAACGCTCGAACTCGGCAATGCCGGCGAACACGGTCAACACCATGCGGCCGGCCGGCGTGGTGGTGTCGGCCCACGGCTCTGCCAGGCTACGCAGGCCCGCGCCGGCCTCCTGGATGCGCTCGGCAATGTCCAGTAGGTCGCGGGTGCTGCGGGCCAGGCGGTCTAGCCTGGTCACTGTCACAACGTCGCCAGGGCGTAGGTGGTCAAGCATCCTGGCCAGCTCCGGGCGGTCGCGCCTGGTGCCGGTGATCTTCTCGGAAAACAGCTTGGTGCAGCCGGCCGCGTGCAGTTCGGCCCGTTGGTTGGTCAAGTCCTGGTCGTCGGTGCTGACGCGGGCATAGCCCAGCAGGCCAGCGGCGGCGCTCTTGTTCATGGCGTAATGTCTCCGGTTCTAGTCGCAAGTATTCTACTTTATGCGACTAAAACACGCGACAAGAAAACGCCAGGAAAAGGGCAGGGCGGCAGCCTGTCGCGTAACTTAGGACTTGTGCGACATGTCGTTTTCAGAAGACGGCTGCACTGAACGTCAGAAGCCGACTGCACTATAGCAGCGGAGGGGTTGGATCAAAGTACTTTAAAGTACTTTAAAGTACTTTAAAGTACTTTGATCCCGAGGGGAACCCTGTGGTTGGCATGCACATACAAATGGACGAACGGATAAACCTTTTCACGCCCTTTTAAATATCCGATTATTCTAATAAACG
next plasmid