pSecTag2 A
Search name
pSecTag2 A,Plasmid pSecTag2 A,pSecTag2 A vector
pSecTag2 A Information
Promoter: CMV
Replicon: pUC ori, FI ori
Terminator: SV40 poly (A) signal, BGH poly (A) signal
Plasmid size: 5159bp
Prokaryotic resistance: ampicillin Amp
Screening marker: bleomycin Zeo
Clone strain: Escherichia coli DH5 alpha
Culture conditions: aerobic at 37 C, LB
Expression host: lactation cells
Induction mode: no need to induce, transient expression.
5'sequencing primers: T7:TAATACGACTCACTATAGGG
3'sequencing primers: BGH:TAGAAGGCACAGTCGAGG
pSecTag2 A Description
pSecTag2 A, B, and C vectors are fusion vectors requiring that you clone your gene of interest in frame with the initiation ATG of the N-terminal Igκ-chain leader sequence and/or the C-terminal myc epitope/polyhistidine tag. Three versions of this vector are provided to facilitate cloning. For proper expression,first determine which restriction sites are appropriate for ligation and then which vector will preserve the reading frame at BOTH the 5′ and the 3′ ends. It may be nece ssary to PCR your gene product to create a fragment with the appropriate restriction sites to clone in frame at both ends. Carefully inspect your gene and the multiple cloning site of each vector before cloning your gene of interest.
pSecTag2 A Multiple cloning site
pSecTag2 A Sequence
LOCUS Exported 5159 bp ds-DNA circular SYN 29-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5159)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, August 29, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..5159
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 905..967
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/note="Ig-kappa leader"
/translation="METDTLLLWVLLLWVPGSTGD"
CDS 1082..1111
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 1127..1144
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1173..1397
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1443..1871
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1885..2214
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2065..2200
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 2262..2309
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 2328..2702
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 2832..2953
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(3002..3018)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3026..3042
/bound_moiety="lac repressor encoded by lacI"
/note="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(3050..3080)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3095..3116
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3404..3992)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4163..5023)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5024..5128)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
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