pBI221-EGFP
Search name
pBI221-EGFP,Plasmid pBI221-EGFP,pBI221-EGFP vector
pBI221-EGFP Information
Promoter: CaMV 35S, Lac
Replicon: ori
Terminator: NOS
Plasmid classification: plant series, protein overexpression vector
Plasmid size: 4530bp
Prokaryotic resistance: Amp
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic LB
5'sequencing primers: 35S:GACGCACAATCCCACTATCC
3'sequencing primers: primers designed according to sequence
Expression: Plant
Use: Plant expression
pBI221-EGFP Description
Most studies related to determining the expression profile of genes and specific promoters used histochemical localization of the reporter gene, gusA. While the histochemical method for visualizing gusA expression suffers from several limitations in the determination of gene expression and location, especially in the tissues with high background acitivty. To solve this problem, a transient expession vector pBI221-RFP/GFP, was constructed using GFP and RFP as double fluorescent marker genes. This vector used CaMV 35S promoter to drive GFP and determine the transforming efficiency. It analyzed expression profile of the target gene and promoter through the RFP activities of the tranformed tissues. Through the specific promoter AGPL1 from watermelon and E8 promoter from tomato, it is resistible to use this vector to study the expression patterns of promoters. Results indicated that the pBI221-RFP/GFP is a very efficient transient expression vector that can be verify the functions of the genes and promoters.
pBI221-EGFP Sequence
LOCUS Exported 4530 bp ds-DNA circular SYN 13-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4530)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 13, 2016 from SnapGene Viewer 3.2.1
FEATURES Location/Qualifiers
source 1..4530
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 512..857
/note="CaMV 35S promoter"
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 903..1622
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
terminator 1641..1893
/note="NOS terminator"
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(1902..1918)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 2392..2496
/gene="bla"
/note="AmpR promoter"
CDS 2497..3357
/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"
rep_origin 3528..4116
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4404..4425
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4440..4470
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4478..4494
/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)."
primer_bind 4502..4518
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 aagcttgcat gcctgcaggt ccccagatta gccttttcaa tttcagaaag aatgctaacc
61 cacagatggt tagagaggct tacgcagcag gtctcatcaa gacgatctac ccgagcaata
121 atctccagga aatcaaatac cttcccaaga aggttaaaga tgcagtcaaa agattcagga
181 ctaactgcat caagaacaca gagaaagata tatttctcaa gatcagaagt actattccag
241 tatggacgat tcaaggcttg cttcacaaac caaggcaagt aatagagatt ggagtctcta
301 aaaaggtagt tcccactgaa tcaaaggcca tggagtcaaa gattcaaata gaggacctaa
361 cagaactcgc cgtaaagact ggcgaacagt tcatacagag tctcttacga ctcaatgaca
421 agaagaaaat cttcgtcaac atggtggagc acgacacact tgtctactcc aaaaatatca
481 aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg gtaatatccg
541 gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag atagtggaaa
601 aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc gttgaagatg
661 cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
721 aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa
781 gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat
841 ttcatttgga gagaacacgg gggactctag aggatccgaa ttcgagctcc gtcgacaagc
901 ttatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
961 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
1021 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
1081 ccctcgtgac caccttcacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
1141 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
1201 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
1261 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
1321 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
1381 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
1441 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
文献和实验
技术资料