質(zhì)粒簡(jiǎn)介
pQE-30Xa質(zhì)粒含有一個(gè)強(qiáng)大的噬菌體T5啟動(dòng)子（被E.coli RNA 聚合酶識(shí)別），并含有兩個(gè)lac操縱子遏制模塊，用于在E.coli中嚴(yán)謹(jǐn)調(diào)控，高水平表達(dá)融合蛋白。在高水平lac遏制子存在的情況下，蛋白合成被有效阻斷，細(xì)胞**載體的穩(wěn)定性增強(qiáng)，pQE載體可以在重組蛋白的N端或C端帶上6×His標(biāo)簽。
質(zhì)粒序列
LOCUS Exported 3509 bp ds-DNA circular SYN 09-JAN-2013
DEFINITION Bacterial vector for expressing N-terminally 6xHis-tagged proteins
with a Factor Xa cleavage site.
ACCESSION .
VERSION .
KEYWORDS pQE-30 Xa
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3509)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
COMMENT Clone at StuI to insert a coding sequence just after the Factor Xa
cleavage site.
FEATURES Location/Qualifiers
source 1..3509
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 10..54
/note="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 101..106
/note="ribosome binding site"
CDS 115..117
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 127..144
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 166..177
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
misc_feature 175..240
/note="MCS"
/note="multiple cloning site"
misc_feature 240..250
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 256..350
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 394..1053
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1118..1204
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(1685..2273)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2444..3304)
/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(3305..3409)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatcgcatc accatcacca tcacggatct ggctctggat ctggtatcga gggaaggcct
181 tataatggaa ctggatccgc atgcgagctc ggtaccccgg gtcgacctgc agccaagctt
241 aattagctga gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
301 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagctagc
361 ttggcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
421 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
481 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
541 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
601 catccggaat ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
661 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
721 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
781 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
841 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
901 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
961 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
1021 cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct
1081 taaacgcctg gggtaatgac tctctagctt gaggcatcaa ataaaacgaa aggctcagtc
1141 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
1201 aaatccgccc tctagagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
1261 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
1321 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
1381 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
1441 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
1501 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
1561 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
1621 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
1681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
1741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
1801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
1861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
1921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
1981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
2041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
2101 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
2161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
2221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
2281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
2341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
2401 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
2461 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
2521 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
2581 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
2641 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
2701 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
2761 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
2821 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
2881 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
2941 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
3001 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
3061 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
3121 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
3181 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
3241 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
3301 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
3361 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
3421 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
3481 ggcgtatcac gaggcccttt cgtcttcac
//