質(zhì)粒簡介
pQE-30質(zhì)粒含有一個強大的噬菌體T5啟動子（被E.coli RNA 聚合酶識別），并含有兩個lac操縱子遏制模塊，用于在E.coli中嚴謹調(diào)控，高水平表達融合蛋白。在高水平lac遏制子存在的情況下，蛋白合成被有效阻斷，細胞**載體的穩(wěn)定性增強，pQE載體可以在重組蛋白的N端或C端帶上6×His標簽。
質(zhì)粒序列
LOCUS Exported 3461 bp ds-DNA circular SYN 08-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3461)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, September 8, 2016 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..3461
/organism="synthetic DNA construct"
/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 97..108
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 127..144
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 208..302
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 346..1005
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1070..1156
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
misc_feature 1311..1451
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(1637..2225)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2396..3256)
/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(3257..3361)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatcgcatc accatcacca tcacggatcc gcatgcgagc tcggtacccc gggtcgacct
181 gcagccaagc ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag
241 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
301 atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
361 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
421 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
481 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
541 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
601 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
661 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
721 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
781 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
841 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
901 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
961 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
1021 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
1081 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
1141 cctgagtagg acaaatccgc cctctagagc tgcctcgcgc gtttcggtga tgacggtgaa
1201 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
1261 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
1321 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
1381 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
1441 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
1501 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
1561 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
1621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
1681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
1741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
1801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
1861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
1921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
1981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
2041 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
2101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
2161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
2221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
2281 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
2341 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
2401 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
2461 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
2521 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
2581 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
2641 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
2701 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
2761 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
2821 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
2881 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
2941 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
3001 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
3061 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
3121 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
3181 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
3241 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
3301 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
3361 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
3421 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//