pX037 vector (V017022)

Price Information

Cat No. Plasmid Name Availability Add to cart
V017022 pX037 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:
pX037
Antibiotic Resistance:
Kanamycin
Length:
19895 bp
Type:
Protein expression
Replication origin:
oriV
Host:
Plants
Selection Marker:
Neo/G418
Promoter:
CaMV 35S
Growth Strain(s):
DH5a
Growth Temperature:
37℃

pX037 vector Map

pX03719895 bp90018002700360045005400630072008100900099001080011700126001350014400153001620017100180001890019800trfALB T-DNA repeatNOS promoterTMV OmegaNeoR/KanROCS terminatorCaMV 35S promoterminimal CaMV 35S promoterTMV OmegaHispidin-3-hydroxylaseLuciferase6xHisOCS terminatorCaMV 35S promoterCAP binding sitelac promoterlac operatorM13 revMCSRB T-DNA repeatoriVAmpR promoterKanR

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.

pX037 vector Sequence

LOCUS       V017022                19895 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017022
VERSION     V017022
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 19895)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..19895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             749..1894
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     misc_feature    2043..2067
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2230..2409
                     /label="NOS promoter"
                     /note="nopaline synthase promoter"
     misc_feature    2440..2494
                     /label="TMV Omega"
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             2496..3287
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     terminator      3317..4022
                     /label="OCS terminator"
                     /note="octopine synthase terminator"
     promoter        4124..4469
                     /label="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     promoter        10772..10818
                     /label="minimal CaMV 35S promoter"
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    10849..10902
                     /label="TMV Omega"
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             10911..12176
                     /gene="h3h"
                     /label="Hispidin-3-hydroxylase"
                     /note="Hispidin-3-hydroxylase from Neonothopanus nambi.
                     Accession#: A0A3G9K5C8"
     CDS             13434..14234
                     /gene="luz"
                     /label="Luciferase"
                     /note="Luciferase from Neonothopanus nambi. Accession#:
                     A0A3G9JYH7"
     CDS             14241..14258
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      14286..14993
                     /label="OCS terminator"
                     /note="octopine synthase terminator"
     promoter        15098..15443
                     /label="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    17242..17263
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        17278..17308
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    17316..17332
                     /label="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)."
     primer_bind     17340..17356
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(17369..17425)
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     misc_feature    17807..17831
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      17918..18629
                     /label="oriV"
                     /note="incP origin of replication"
     promoter        18650..18754
                     /label="AmpR promoter"
     CDS             18755..19546
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"