pLSLZ vector (V011387)

Price Information

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V011387 pLSLZ In stock, 1 week for quality controls

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

Basic Vector Information

Vector Name:
pLSLZ
Antibiotic Resistance:
Kanamycin
Length:
15655 bp
Type:
Plant Expression Vectors
Replication origin:
ori
Host:
Plants
Promoter:
CaMV 35S (enhanced)

pLSLZ vector Map

pLSLZ15655 bp70014002100280035004200490056006300700077008400910098001050011200119001260013300140001470015400attB1SV40 NLSEarly growth response protein 1FokI cleavage domainNOS terminatorattB5NeoR/KanRNOS terminatorattB2CaMV 35S promoterM13 revlac operatorlac promoterCAP binding siteCaMV 35S promoter (enhanced)HygRCaMV poly(A) signalLB T-DNA repeatKanRoribompVS1 oriVpVS1 RepApVS1 StaARB T-DNA repeatM13 fwd

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.

pLSLZ vector Sequence

LOCUS       V011387                15655 bp    DNA     circular SYN 13-JAN-2022
DEFINITION  Exported.
ACCESSION   V011387
VERSION     V011387
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 15655)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 15655)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..15655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    529..553
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             587..607
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             641..817
                     /gene="EGR1"
                     /label="Early growth response protein 1"
                     /note="Early growth response protein 1 from Serinus
                     canaria. Accession#: O73694"
     CDS             875..1462
                     /label="FokI cleavage domain"
                     /note="nonspecific DNA cleavage domain of the FokI
                     endonuclease (Li et al., 1992)"
     terminator      1476..1728
                     /label="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    1773..1793
                     /label="attB5"
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     CDS             2824..3612
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     terminator      3642..3894
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    complement(4907..4931)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5697..6041
                     /label="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(6752..6768)
                     /label="M13 rev"
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    6776..6792
                     /label="lac repressor encoded by lacI binding site"
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6800..6830)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6845..6866)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7057..7734
                     /label="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     CDS             7802..8824
                     /label="HygR"
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    8868..9042
                     /label="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    complement(9120..9144)
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9569..10360
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      10450..11038
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(11224..11364)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(11708..11902)
                     /direction=LEFT
                     /label="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11971..13035)
                     /label="pVS1 RepA"
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13472..14098)
                     /label="pVS1 StaA"
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     misc_feature    complement(15398..15422)
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15625..15641
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"