pTYB21 vector (V012374)

Basic Vector Information

      • Vector Name:
      • pTYB21
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7514 bp
      • Type:
      • Cloning Vectors
      • Source/Author:
      • Chong S, Montello GE, Zhang A, Cantor EJ, Liao W, Xu MQ, Benner J.
      • Copy Number:
      • High copy number

pTYB21 vector Vector Map

pTYB217514 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500AmpR promoterAmpRM13 orioriropCAP binding sitelacIlacI promoterrrnB T1 terminatorrrnB T1 terminatorrrnB T1 terminatorrrnB T1 terminatorrrnB T1 terminatorT7 promoterlac operatorRBSMBP peptideSce VMA intein 5' regionCBDSce VMA intein 3' regionMCST7 terminator

Plasmid Resuspension 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 ℃.

pTYB21 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pTYB21.        7514 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial expression vector that encodes a thiol-cleavable 
            N-terminal intein with a chitin-binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    pTYB21.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Chong S, Montello GE, Zhang A, Cantor EJ, Liao W, Xu MQ, Benner J.
  TITLE     Utilizing the C-terminal cleavage activity of a protein splicing 
            element to purify recombinant proteins in a single chromatographic 
            step.
  JOURNAL   Nucleic Acids Res. 1998;26:5109-15.
  PUBMED    9801307
REFERENCE   2  (bases 1 to 7514)
  AUTHORS   New England Biolabs
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7514)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "1998"; volume: "26"; pages: "5109-15"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Clone with the SapI site to generate a fusion protein that can be 
            isolated with no added amino acids.
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..139
                     /label=AmpR promoter
     CDS             140..997
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      complement(1042..1555)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      1666..2254
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2626..2814)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3337..3358)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3374..4453)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4454..4531)
                     /label=lacI promoter
     terminator      4684..4770
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4867..4953
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5050..5136
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5233..5319
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(5419..5462)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        5637..5655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5656..5680
                     /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)."
     RBS             5695..5717
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5725..5754
                     /codon_start=1
                     /product="N-terminal peptide from mature maltose-binding
                     protein"
                     /label=N-terminal peptide from mature maltose-binding
                     /note="MBP peptide"
                     /translation="MKIEEGKLVI"
     CDS             5770..6588
                     /label=Sce VMA intein 5' region
                     /note="5' region of the intein from the yeast Vma1 subunit
                     of the vacuolar ATPase (Chong et al., 1998)"
     CDS             6592..6747
                     /label=CBD
                     /note="chitin binding domain from chitinase A1 (Watanabe et
                     al., 1994)"
     CDS             6751..7299
                     /label=Sce VMA intein 3' region
                     /note="modified 3' region of the intein from the yeast Vma1
                     subunit of the vacuolar ATPase (Chong et al., 1998)"
     misc_feature    7300..7377
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      7442..7489
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"

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