pSecTag2 A Plasmid

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  • Model: PVT1040
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pSecTag2 A

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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 vector

 

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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