pTYB21 vector (V012374)

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"