pT7-His-pCSK9
Catalog No. PVT10138
Packing 2ug
pT7-His-pCSK9 Information
Function ORF expression plasmid
Resistance Kan
Screen /
Strain DH5alpha
Culture temperature 37degrees centigrade
Replicon
Copy
Promoter
Induction
Forward primer
Reverse primer
pT7-His-pCSK9 Description
PT7-His-pCSK9 is a human gene expression plasmid for large intestine.
pT7-His-pCSK9 Sequence
LOCUS Exported 7366 bp ds-DNA circular SYN 24-Aug-2017
FEATURES Location/Qualifiers
source 1..7366
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 141..159
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 160..184
/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)."
misc_feature 199..221
/note="RBS"
CDS 240..257
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 282..2360
/codon_start=1
/note="pCSK9"
/translation="MGTVSSRRSWWPLPLLLLLLLLLGPAGARAQEDEDGDYEELVLAL
RSEEDGLAEAPEHGTTATFHRCAKDPWRLPGTYVVVLKEETHLSQSERTARRLQAQAAR
RGYLTKILHVFHGLLPGFLVKMSGDLLELALKLPHVDYIEEDSSVFAQSIPWNLERITP
PRYRADEYQPPDGGSLVEVYLLDTSIQSDHREIEGRVMVTDFENVPEEDGTRFHRQASK
CDSHGTHLAGVVSGRDAGVAKGASMRSLRVLNCQGKGTVSGTLIGLEFIRKSQLVQPVG
PLVVLLPLAGGYSRVLNAACQRLARAGVVLVTAAGNFRDDACLYSPASAPEVITVGATN
AQDQPVTLGTLGTNFGRCVDLFAPGEDIIGASSDCSTCFVSQSGTSQAAAHVAGIAAMM
LSAEPELTLAELRQRLIHFSAKDVINEAWFPEDQRVLTPNLVAALPPSTHGAGWQLFCR
TVWSAHSGPTRMATAVARCAPDEELLSCSSFSRSGKRRGERMEAQGGKLVCRAHNAFGG
EGVYAIARCCLLPQANCSVHTAPPAEASMGTRVHCHQQGHVLTGCSSHWEVEDLGTHKP
PVLRPRGQPNQCVGHREASIHASCCHAPGLECKVKEHGIPAPQEQVTVACEEGWTLTGC
SALPGTSHVLGAYAVDNTCVVRSRDVSTTGSTSEGAVTAVAICCRSRHLAQASQELQ"
CDS 2367..2384
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 2451..2498
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2535..2990
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(3083..3898)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 4020..4608
/direction=RIGHT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5038..5229)
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6038..7120)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="lacI"
/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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(7121..7198)
/gene="lacI"
/note="lacI promoter"
ORIGIN
1 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
61 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
121 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
181 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggcagcagcc
241 atcatcatca tcatcacagc agcggccgcg gatccgaatt catgggcacc gtcagctcca
301 ggcggtcctg gtggccgctg ccactgctgc tgctgctgct gctgctcctg ggtcccgcgg
361 gcgcccgtgc gcaggaggac gaggacggcg actacgagga gctggtgcta gccttgcgtt
421 ccgaggagga cggcctggcc gaagcacccg agcacggaac cacagccacc ttccaccgct
481 gcgccaagga tccgtggagg ttgcctggca cctacgtggt ggtgctgaag gaggagaccc
文献和实验
技术资料