Basic Vector Information
- Vector Name:
- pAT224
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5189 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Binz HK, Amstutz P, Kohl A, Stumpp MT, Briand C, Forrer P, Grutter MG, Pluckthun A.
- Promoter:
- T5
pAT224 vector Map
pAT224 vector Sequence
LOCUS 40924_4974 5189 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pAT224, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5189)
AUTHORS Binz HK, Amstutz P, Kohl A, Stumpp MT, Briand C, Forrer P, Grutter
MG, Pluckthun A.
TITLE High-affinity binders selected from designed ankyrin repeat protein
libraries
JOURNAL Nat. Biotechnol. 22 (5), 575-582 (2004)
PUBMED 15097997
REFERENCE 2 (bases 1 to 5189)
AUTHORS Stumpp MT, Forrer P, Binz KH, Amstutz P, Zahnd C, Pluckthun A.
TITLE Direct Submission
JOURNAL Submitted (20-JUN-2003) Biochemistry, University of Zurich,
Winterthurerstrasse 190, Zurich 8057, Switzerland
REFERENCE 3 (bases 1 to 5189)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5189)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Biotechnol."; date: "2004"; volume: "22"; issue: "5"; pages:
"575-582"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(20-JUN-2003) Biochemistry, University of Zurich,
Winterthurerstrasse 190, Zurich 8057, Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5189
/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"
CDS 178..1275
/label=MBP
/note="maltose binding protein from E. coli"
misc_feature 1276..1317
/label=multiple cloning site
/note="multiple cloning site"
CDS 1318..1335
/label=6xHis
/note="6xHis affinity tag"
terminator 1352..1446
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator 1553..1639
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1702..2781)
/label=lacI
/note="lac repressor"
promoter complement(2782..2859)
/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 3039..3179
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3365..3953)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4127..4984)
/label=AmpR
/note="beta-lactamase"
promoter complement(4985..5089)
/label=AmpR promoter
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