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pYPQ141-ZmUbi-RZ-Lb vector (V005899)

Price Information

Cat No. Plasmid Name Availability Add to cart
V005899 pYPQ141-ZmUbi-RZ-Lb In stock, 1 week for quality controls

Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pYPQ141-ZmUbi-RZ-Lb
Antibiotic Resistance:
Spectinomycin
Length:
4991 bp
Type:
CRISPR
Replication origin:
ori
Copy Number:
High Copy
Promoter:
Maize ubiquitin 1
Cloning Method:
Gateway Cloning
5' Primer:
ttcccagtcacgacgttgtaaaac
3' Primer:
catggtcatagctgtttcctg

pYPQ141-ZmUbi-RZ-Lb vector Map

Copy Sequence View Map
pYPQ141-ZmUbi-RZ-Lb4991 bp6001200180024003000360042004800hammerhead ribozymeHDV ribozymeattL2T7 promoterT7 promoterM13 revSmRoriL4440rrnB T2 terminatorrrnB T1 terminatorM13 fwdattL5Ubi promoter

Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pYPQ141-ZmUbi-RZ-Lb vector Sequence

LOCUS       40924_48233        4991 bp DNA     circular SYN 13-MAY-2021
DEFINITION  LbCpf1 Gateway gRNA entry plasmid using Zea mays Ubi promoter and 
            ribozyme processing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  AUTHORS   Tang X, Lowder LG, Zhang T, Malzahn AA, Zheng X, Voytas DF, Zhong Z,
            Chen Y, Ren Q, Li Q, Kirkland ER, Zhang Y, Qi Y
  TITLE     A CRISPR-Cpf1 system for efficient genome editing and 
            transcriptional repression in plants.
  JOURNAL   Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18.
  PUBMED    28211909
REFERENCE   2  (bases 1 to 4991)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4991)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Plants."; date: "2017-02-17"; pages: "
            10.1038/nplants.2017.18"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     ncRNA           1040..1076
                     /label=hammerhead ribozyme
     ncRNA           1130..1197
                     /label=HDV ribozyme
     protein_bind    complement(1217..1316)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1334..1352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1369..1387)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1392..1408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1839..2627
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2723..3311
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3465..3482
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3641..3668)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3760..3846)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     3910..3926
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3941..4036
                     /label=attL5
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4048..4991
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"