pHANNIBAL vector (V010843)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V010843	pHANNIBAL	In stock, instant shipping	Buy one, get one free!

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

Basic Vector Information

Vector Name:: pHANNIBAL

Antibiotic Resistance:: Ampicillin

Length:: 5824 bp

Type:: Plant Vectors

Replication origin:: ori

Host:: Plants

Source/Author:: Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding

Copy Number:: High copy number

Promoter:: CaMV 35S

pHANNIBAL vector Vector Map

View Map in NovoBuilder

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.

pHANNIBAL vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pHANNIBAL.        5824 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for silencing genes in plants using intron-containing hairpin
            RNA (hpRNA).
ACCESSION   .
VERSION     .
KEYWORDS    pHANNIBAL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  AUTHORS   Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, Liu Q, Gooding
            PS, Singh SP, Abbott D, Stoutjesdijk PA, Robinson SP, Gleave AP, 
            Green AG, Waterhouse PM.
  TITLE     Construct design for efficient, effective and high-throughput gene 
            silencing in plants.
  JOURNAL   Plant J. 2001;27:581-90.
  PUBMED    11576441
REFERENCE   2  (bases 1 to 5824)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5824)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant J."; 
            date: "2001"; volume: "27"; pages: "581-90"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..35)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(215..643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        670..774
                     /label=AmpR promoter
     CDS             775..1632
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1806..2394
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2682..2703
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2718..2748
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2756..2772
                     /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     2780..2796
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2827..2845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3863..4208
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    4210..4228
                     /label=MCS 1
                     /note="MCS 1"
                     /note="multiple cloning site 1"
     intron          4236..5002
                     /label=PDK intron
                     /note="pyruvate orthophosphate dikinase intron from
                     Flaveria trinervia"
     misc_feature    5022..5045
                     /label=MCS 2
                     /note="MCS 2"
                     /note="multiple cloning site 2"
     terminator      5049..5755
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(5796..5814)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"