pComb3X is a new version of the pComb vectors. Improvements over pComb3 include increased stability and the introduction of an asymmetric SfiI cassette for directional cloning of full Fab, scFv, peptide, and other proteins for phage display. 6xHis and HA tags allow for purification and detection. An amber stop codon was introduced to turn off the expression of the pIII fusion protein by switching to a non-suppressor strain of E. coli allowing the production of soluble protein without subcloning.
| 载体基本信息 | |||
| 载体名称 | pComb3X | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 3451 bp | 载体类型 | Phagemid cloning vector |
| 载体来源 | Rader C, Barbas CF III. | ||
pComb3X 载体载体图谱
pComb3X 载体载体序列
LOCUS Exported 3451 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Phagemid cloning vector pComb3X, complete sequence.
ACCESSION AF268281
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3451)
AUTHORS Rader C, Barbas CF III.
JOURNAL (in) PHAGE DISPLAY, A LABORATORY MANUAL. Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, NY, USA (2000), In press
REFERENCE 2 (bases 1 to 3451)
AUTHORS Rader C, Barbas CF III.
TITLE Direct Submission
JOURNAL Submitted (16-MAY-2000) Department of Molecular Biology, BCC-526,
Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA
92037, USA
REFERENCE 3 (bases 1 to 3451)
AUTHORS Rader C, Fuller R, Barbas CF III.
TITLE Direct Submission
JOURNAL Submitted (05-SEP-2003) Department of Molecular Biology, BCC-526,
Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA
92037, USA
REFERENCE 4 (bases 1 to 3451)
AUTHORS .
TITLE Direct Submission
COMMENT On Sep 5, 2003 this sequence version replaced AF268281.1.
FEATURES Location/Qualifiers
source 1..3451
/organism="Phagemid cloning vector pComb3X"
/mol_type="genomic DNA"
/db_xref="taxon:137786"
protein_bind 109..130
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 145..175
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 183..199
/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)."
CDS 329..394
/codon_start=1
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/label=pelB signal sequence
/translation="MKYLLPTAAAGLLLLAAQPAMA"
CDS 440..457
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 464..490
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
rep_origin complement(1066..1521)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1548..1652
/gene="bla"
/label=AmpR promoter
CDS 1653..2513
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2684..3272
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
