pRSET-BFP
Search name
pRSET-BFP,Plasmid pRSET-BFP,pRSET-BFP vector
pRSET-BFP Information
Promoter: T7
Replicator: pUC ori, F1 ori
Terminator: T7 Terminator
Plasmid classification: Escherichia coli vector; pRSET series expression plasmid
Plasmid size: 3600bp
Plasmid label: N-His, N-EK, N-Xpress
Prokaryotic resistance: ampicillin Amp
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Express host: BL21 (DE3) series, recommend BL21Star (DE3)
Culture conditions: 37 centigrade, aerobic, LB
Inducement: IPTG or lactose and its analogues
5'sequencing primers: T7 Forward: 5-TAATACGACTCACTATAGGG-3
3'sequencing primers: Based on sequence design
Remarks: pRSET expression plasmid
pRSET-BFP Description
pRSET-BFP Sequence
LOCUS Exported 3600 bp ds-DNA circular SYN 29-七月-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pRSET-BFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3600)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2016-7-28 from SnapGene Viewer 2.8.1
FEATURES Location/Qualifiers
source 1..3600
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 9..27
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 101..118
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 122..154
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 158..181
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/note="Xpress(TM) tag"
/translation="DLYDDDDK"
CDS 209..928
/codon_start=1
/product="blue variant of GFP"
/note="BFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
terminator 1004..1051
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1148..1603
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1629..1733
/gene="bla"
/note="AmpR promoter"
CDS 1734..2594
/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 2765..3353
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gcggggttct catcatcatc atcatcatgg
121 tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa
181 ggatcgatgg ggatccgaat tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
241 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
301 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
361 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagccacg gcgtccagtg
421 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
481 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
541 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
601 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacgt
661 ctatatcatg gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
721 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
781 cggccccgtg ctgctgcccg acagccacta cctgagcacc cagtccgccc tgagcaaaga
841 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
901 tctcggcatg gacgagctgt acaagtaact cgagaagctt gatccggctg ctaacaaagc
961 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
1021 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg
1081 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
1141 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
1201 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
1261 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
1321 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
1381 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
1441 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
