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pCAGGS- HA
货源充足
LSM Bio
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2ug
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pCAGGS-HA Information
Promoter: CAG promoter
Replicator: SV40 ori, ori
Plasmid classification: lactation serial plasmid; lactation expression plasmid; pCAG series plasmid.
Plasmid size: 4807bp
Prokaryotic resistance: ampicillin Amp (100 u g/ml)
Cloned strains of Escherichia coli, DH5 A and other Escherichia coli
Culture conditions: 37 centigrade, aerobic, LB
Expression host: mammalian cells such as 293T
Culture conditions: 37 C, 5%CO2
Induction mode: no induction
5'sequencing primers: pcaggs-F (GCTAACCATGTTCATGCCTTCT)
3'sequencing primers: pcaggs-R (TATGTCCTTCCGAGTGAGAG)
pCAGGS-HA Description
pCAGGS-HA is added with HA tag on the basis of pCAGGS vector, which is helpful to detect antibody in WB test. pCAGGS-HA plasmid is useful for highly efficient expression of genes under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.The AG promoter sequence consists of the chicken β-actin promoter, the first exon and part of the first intron (that seems to have a strong enhancer-like activity) linked to a rabbit β-globin fragment, consisting of a 3' part of the second intron (inclusive a branch point which is required for normal splicing reactions) and a 5' part of the third exon.When cloning a fragment downstream from the lac promoter it may be advisable to use lacIq strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
pCAGGS-HA Sequence
LOCUS Exported 4807 bp ds-DNA circular SYN 17-JUL-2016
DEFINITION synthetic circular DNA
KEYWORDS pCAGGS-HA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4807)
TITLE Direct Submission
JOURNAL Exported Sunday, July 17, 2016
FEATURES Location/Qualifiers
source 1..4807
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 385..662
/note="chicken beta-actin promoter"
intron 663..1680
/note="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
misc_feature 1735..1737
/note="M"
CDS 1738..1764
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
polyA_site 1894..1949
/note="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(2310..2326)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2334..2350
/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(2358..2388)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2403..2424
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2483..2678
/note="SV40 promoter"
/note="SV40 early promoter"
rep_origin 2529..2664
/note="SV40 ori"
/note="SV40 origin of replication"
polyA_signal 2684..2818
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(3056..3644)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3815..4675)
/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(4676..4780)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
481 cgatgggggc gggggggggg ggggggcccc ccccaggcgg ggcggggcgg ggcgaggggc
541 ggggcggggc gaggcggaaa ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
661 cgggagtcgt tgcgcgctgc cttccccccg tgccccgctc cgccgccgcc tcgcgccgcc
721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
1261 cggggccgcc tcgggccggg gagggctcgg gggaaggggc gcggcggccc ccggagcgcc
1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
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