pAT223 vector (V009414)

Basic Vector Information

Vector Name:
pAT223
Antibiotic Resistance:
Ampicillin
Length:
4366 bp
Type:
Expression vector
Replication origin:
ori
Source/Author:
Amstutz P, Binz HK, Parizek P, Stumpp MT, Kohl A, Grutter MG, Forrer P, Pluckthun A.

pAT223 vector Map

pAT2234366 bp600120018002400300036004200T5 promoterlac operatorRBSAviTag(TM)bacteriophage lambda protein D6xHismultiple cloning sitelambda t0 terminatorrrnB T1 terminatorlacIlacIq promoterbomoriAmpRAmpR promoter

pAT223 vector Sequence

LOCUS       40924_4969        4366 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pAT223, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4366)
  AUTHORS   Amstutz P, Binz HK, Parizek P, Stumpp MT, Kohl A, Grutter MG, Forrer
            P, Pluckthun A.
  TITLE     Intracellular kinase inhibitors selected from combinatorial 
            libraries of designed ankyrin repeat proteins
  JOURNAL   J. Biol. Chem. 280 (26), 24715-24722 (2005)
  PUBMED    15851475
REFERENCE   2  (bases 1 to 4366)
  AUTHORS   Stumpp MT, Forrer P, Binz HK, Amstutz P, Zahnd C, Pluckthun A.
  TITLE     Designed ankyrin repeat proteins binding maltose binding protein
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4366)
  AUTHORS   Stumpp MT, Forrer P, Binz HK, Amstutz P, Zahnd C, Pluckthun A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland
REFERENCE   4  (bases 1 to 4366)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4366)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Biol. 
            Chem."; date: "2005"; volume: "280"; issue: "26"; pages: 
            "24715-24722"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    62..78
                     /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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             121..165
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
     misc_feature    172..456
                     /label=bacteriophage lambda protein D
                     /note="bacteriophage lambda protein D"
     CDS             457..474
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    475..519
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      529..623
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      730..816
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(879..1958)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1959..2036)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    2216..2356
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2542..3130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3304..4161)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4162..4266)
                     /label=AmpR promoter

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