Basic Vector Information
pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins. Each vector has a large multiple cloning site in three reading frames to simplify cloning in frame with the N-terminal secretion signal. The vectors (Figure 1) offer the following features:1.Secretion signal from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins; 2. Cytomegalovirus (CMV) promoter for high-level constitutive expression; 3.C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody; 4. C-terminal c-myc epitope for detection with an Anti-myc Antibody; 5. Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability; 5. SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1, COS-7). The pSecTag2 vectors carry the Zeocin™ resistance gene for cost-effective selection in mammalian cells. Zeocin™ selection can also be used in E. coli.The pSecTag2/Hygro vectors have the Hygromycin B resistance gene for selection of stable mammalian cell lines.
- Vector Name:
- pSecTag2/Hygro A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5745 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Thermo Fisher Scientific
- Selection Marker:
- Hygromycin
- Copy Number:
- High copy number
- Promoter:
- CMV
- Cloning Method:
- Enzyme Cut
- 5' Primer:
- TAATACGACTCACTATAGGG
- Fusion Tag:
- His Tag (6x), c-Myc Epitope Tag, IgK Leader Sequence
pSecTag2/Hygro A vector Vector Map
pSecTag2/Hygro A vector Sequence
LOCUS 40924_39143 5745 bp DNA circular SYN 13-DEC-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5745)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5745)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5745
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
sig_peptide 905..967
/label=Ig-kappa leader
/note="leader sequence from mouse immunoglobulin kappa
light
chain"
CDS 1082..1111
/codon_start=1
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
/translation="EQKLISEEDL"
CDS 1127..1144
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
polyA_signal 1173..1397
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1443..1871
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1885..2214
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 2263..3285
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 3418..3551
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3588..3604)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3612..3628)
/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)."
promoter complement(3636..3666)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3681..3702)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3990..4578)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4752..5609)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5610..5714)
/label=AmpR promoter
This page is informational only.