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pBI121 vector (Cat. No.: V010909)

pBI12114758 bp700140021002800350042004900560063007000770084009100980010500112001190012600133001400014700oriVCAP binding sitelac promoterlac operatorM13 revCaMV 35S promoterGUSNOSM13 fwdLB T-DNA repeatsimilar to tetA in GenBank Accession Number L13842trfAKanRIS1similarity to NPT III gene in GenBank Accession Number V01547similar to kilA in GenBank Accession Number M62846oriVtraJoriTColE1 ori; similar to sequence in GenBank Accession Number V00268TetRRB T-DNA repeatNOS promoterNeoR/KanR
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Basic Information

Note: The pBI121 is a binary Agrobacterium vector equipped with a GUS reporter gene designed for facilitating plant transformation.

Name:
pBI121
Antibiotic Resistance:
Kanamycin
Length:
14758 bp
Type:
Plant Vectors
Replication origin:
oriV
Host:
Plants
Source/Author:
Chen PY, Wang CK, Soong SC, To KY.
Selection Marker:
Neomycin/G418(Geneticin)
Copy Number:
Low copy number
Promoter:
CaMV35S(long)
5' Primer:
M13 fwd
3' Primer:
M13 rev
Growth Strain(s):
stbl3
Growth Temperature:
37℃
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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Resources

Plasmid Protocol

Plasmid Reconstitution Protocol

  1. Centrifuge at 5,000×g for 5 min.
  2. Carefully open the tube and add sterile water to dissolve the DNA: add 20 μl for 5 μg plasmid, and 100 μl for 100 μg plasmid.
  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.

References

  • Srinivas K, Muralikrishna N, Kumar KB, Raghu E, Mahender A, Kiranmayee K, Yashodahara V, Sadanandam A. Biolistic transformation of Scoparia dulcis L. Physiol Mol Biol Plants. 2016 Jan;22(1):61-8. doi: 10.1007/s12298-016-0338-2. Epub 2016 Jan 20. PMID: 27186019; PMCID: PMC4840147.

  • Beihaghi M, Marashi H, Bagheri A, Sankian M. Transient expression of CCL21as recombinant protein in tomato. Biotechnol Rep (Amst). 2017 Nov 26;17:10-15. doi: 10.1016/j.btre.2017.11.007. PMID: 29276695; PMCID: PMC5730375.

pBI121 vector (Cat. No.: V010909) Annotation

LOCUS       Exported               14758 bp DNA     circular SYN 30-SEP-2025
DEFINITION  Binary vector pBI121, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14758)
  AUTHORS   Chen PY, Wang CK, Soong SC, To KY.
  TITLE     Complete sequence of the binary vector pBI121 and its application in
            cloning T-DNA insertion from transgenic plants
  JOURNAL   Mol. Breed. 11, 287-293 (2003)
REFERENCE   2  (bases 1 to 14758)
  AUTHORS   To KY.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-FEB-2002) Institute of BioAgricultural Sciences, 
            Academia Sinica, Taipei 11529, Taiwan
REFERENCE   3  (bases 1 to 14758)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 14758)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 14758)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 14758)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Breed.
            11, 287-293 (2003)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (20-FEB-2002) Institute of BioAgricultural Sciences, Academia
            Sinica, Taipei 11529, Taiwan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(10985..14758,1..10984)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(361..986)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     protein_bind    1049..1070
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1085..1115
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1123..1139
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     1147..1163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1687..2032
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2071..3879
                     /label=GUS
                     /note="beta-glucuronidase"
     regulatory      3953..4205
                     /label=NOS
                     /note="NOS"
                     /regulatory_class="terminator"
     terminator      3953..4205
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4214..4230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(4848..4872)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(5382..6424)
                     /label=similar to tetA in GenBank Accession Number L13842
                     /note="similar to tetA in GenBank Accession Number L13842"
     CDS             complement(6471..7616)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(7918..8709)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     mobile_element  8901..9668
                     /label=IS1
                     /note="prokaryotic transposable element"
     misc_feature    complement(9670..10020)
                     /note="similarity to NPT III gene in GenBank Accession
                     Number V01547"
     misc_feature    complement(10021..10292)
                     /label=similar to kilA in GenBank Accession Number M62846
                     /note="similar to kilA in GenBank Accession Number M62846"
     rep_origin      complement(10369..10979)
                     /direction=LEFT
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     CDS             complement(11132..11500)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(11533..11642)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(11774..12152)
                     /direction=LEFT
                     /note="ColE1 ori; similar to sequence in GenBank Accession 
                     Number V00268"
     CDS             complement(12326..12973)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     misc_feature    complement(13438..13462)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        13618..13801
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             13825..14613
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"