pCAGGS
Search name
pCAGGS,Plasmid pCAGGS,pCAGGS vector
pCAGGS Information
Promoter: CAG promoter
Replicator: SV40 ori, ori
Terminator: beta -globin poly (A) signal
Plasmid classification: lactation serial plasmid; lactation expression plasmid; pCAG series plasmid.
Plasmid size: 4801bp
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, instantaneous expression
5'sequencing primers: pcaggs-F (GTTCGGCTTCTGGCGTGT)
3'sequencing primers: pcaggs-R (TATGTCCTTCCGAGTGAGAG)
pCAGGS Description
pCAGGS plasmid can be used to express gene efficiently under the control of chicken b- actin, rabbit b- globulin heterozygous promoter (CAG), and human CMV-IE enhancer in various mammalian cells. The CAG promoter sequence is part of the chicken b- actin promoter, the first exon and the first intron (it seems to have strong enhanced subtype activity. " It is linked to the rabbit b- globin fragment, the 3'part of the second intron includes the branching point needed for normal splicing reaction and the 5' portion of the third exon.
This 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 Sequence
LOCUS Exported 4801 bp ds-DNA circular SYN 18-MAY-2017
DEFINITION synthetic circular DNA
KEYWORDS pCAGGS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4801)
TITLE Direct Submission
JOURNAL Exported Thursday, May 18, 2017
FEATURES Location/Qualifiers
source 1..4801
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_site 158..213
/note="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(574..590)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 598..614
/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(622..652)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 666..687
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 746..941
/note="SV40 promoter"
/note="SV40 early promoter"
rep_origin 792..927
/note="SV40 ori"
/note="SV40 origin of replication"
polyA_signal 947..1081
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(1319..1907)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2078..2938)
/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(2939..3043)
/gene="bla"
/note="AmpR promoter"
enhancer 3074..3453
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3455..3732
/note="chicken beta-actin promoter"
intron 3733..4750
/note="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
ORIGIN
1 ttcctcgagg aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
61 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
121 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
181 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
241 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
301 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
361 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
421 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
481 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
541 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
601 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggt
661 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
721 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
781 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
841 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
901 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
1021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
1081 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
1141 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
1201 gctcactcaa aggcggtaat acggttatcc acagaatcag ggataacgca ggaaagaaca
1261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
1321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
1381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
1441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
1501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
1561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcct <
文献和实验
技术资料