| 质粒名称: | pQE 80L Kan T7WT |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 102790 |
| 存储实验室: | Andrew Ellington |
|
载体骨架基本信息 |
|
| 载体名称: | pQE |
| 载体出品公司: | Qiagen |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 7154 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | T7 RNA polymerase |
| 物种来源: | T7 phage |
| 插入大小: | 2685 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pQE 80L Kan T7WT was a gift from Andrew Ellington (Addgene plasmid # 102790 ; http://n2t.net/addgene:102790 ; RRID:Addgene_102790) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pQE 80L Kan T7WT 7154 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses T7 RNA polymerase.
ACCESSION .
VERSION .
KEYWORDS pQE 80L Kan T7WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7154)
TITLE Compartmentalized Partnered Replication for the Directed Evolution
of Genetic Parts and Circuits
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7154)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7154
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 968..987
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(1007..1084)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
primer_bind complement(1168..1190)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 1276..1416
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 1331..1350
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(1431..1448)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1602..2190)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1682..1701)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2297..3112)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 3020..3039
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3104..3123
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind 3184..3202
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3226..3270
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 3246..3262
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
protein_bind 3278..3294
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
RBS 3313..3324
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 3343..3360
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 6037..6131
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 6175..6834
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(6241..6260)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 6899..6985
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(7078..1006)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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"
ORIGIN
1 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
61 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
121 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
181 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
241 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
301 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
361 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
421 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
481 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
541 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
601 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
661 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
721 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
781 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
841 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
901 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
961 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
1021 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac cattcgatgg
1081 tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg cgcatgatct agagctgcct
1141 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
1201 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
1261 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
1321 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
1381 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
1441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
1501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
1561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
1621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
1681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
1741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
1801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
1861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
1921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
1981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
2041 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
2101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
2161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
2221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggcgtt aagggatttt
2281 ggtcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
2341 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
2401 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
2461 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
2521 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
2581 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
2641 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
2701 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
2761 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
2821 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
2881 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
2941 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
3001 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
3061 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
3121 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccatgac attaacctat
3181 aaaaataggc gtatcacgag gccctttcgt cttcacctcg agaaatcata aaaaatttat
3241 ttgctttgtg agcggataac aattataata gattcaattg tgagcggata acaatttcac
3301 acagaattca ttaaagagga gaaattaact atgagaggat cgcatcacca tcaccatcac
3361 ggatccaaca cgattaacat cgctaagaac gacttctctg acatcgaact ggctgctatc
3421 ccgttcaaca ctctggctga ccattacggt gagcgtttag ctcgcgaaca gttggccctt
3481 gagcatgagt cttacgagat gggtgaagca cgcttccgca agatgtttga gcgtcaactt
3541 aaagctggtg aggttgcgga taacgctgcc gccaagcctc tcatcactac cctactccct
3601 aagatgattg cacgcatcaa cgactggttt gaggaagtga aagctaagcg cggcaagcgc
3661 ccgacagcct tccagttcct gcaagaaatc aagccggaag ccgtagcgta catcaccatt
3721 aagaccactc tggcttgcct aaccagtgct gacaatacaa ccgttcaggc tgtagcaagc
3781 gcaatcggtc gggccattga ggacgaggct cgcttcggtc gtatccgtga ccttgaagct
3841 aagcacttca agaaaaacgt tgaggaacaa ctcaacaagc gcgtagggca cgtctacaag
3901 aaagcattta tgcaagttgt cgaggctgac atgctctcta agggtctact cggtggcgag
3961 gcgtggtctt cgtggcataa ggaagactct attcatgtag gagtacgctg catcgagatg
4021 ctcattgagt caaccggaat ggttagctta caccgccaaa atgctggcgt agtaggtcaa
4081 gactctgaga ctatcgaact cgcacctgaa tacgctgagg ctatcgcaac ccgtgcaggt
4141 gcgctggctg gcatctctcc gatgttccaa ccttgcgtag ttcctcctaa gccgtggact
4201 ggcattactg gtggtggcta ttgggctaac ggtcgtcgtc ctctggcgct ggtgcgtact
4261 cacagtaaga aagcactgat gcgctacgaa gacgtttaca tgcctgaggt gtacaaagcg
4321 attaacattg cgcaaaacac cgcatggaaa atcaacaaga aagtcctagc ggtcgccaac
4381 gtaatcacca agtggaagca ttgtccggtc gaggacatcc ctgcgattga gcgtgaagaa
4441 ctcccgatga aaccggaaga catcgacatg aatcctgagg ctctcaccgc gtggaaacgt
4501 gctgccgctg ctgtgtaccg caaggacaag gctcgcaagt ctcgccgtat cagccttgag
4561 ttcatgcttg agcaagccaa taagtttgct aaccataagg ccatctggtt cccttacaac
4621 atggactggc gcggtcgtgt ttacgctgtg tcaatgttca acccgcaagg taacgatatg
4681 accaaaggac tgcttacgct ggcgaaaggt aaaccaatcg gtaaggaagg ttactactgg
4741 ctgaaaatcc acggtgcaaa ctgtgcgggt gtcgataagg ttccgttccc tgagcgcatc
4801 aagttcattg aggaaaacca cgagaacatc atggcttgcg ctaagtctcc actggagaac
4861 acttggtggg ctgagcaaga ttctccgttc tgcttccttg cattctgctt tgagtacgct
4921 ggggtacagc accacggcct gagctataac tgctcccttc cgctggcgtt tgacgggtct
4981 tgctctggca tccagcactt ctccgcgatg ctccgagatg aggtaggtgg tcgcgcggtt
5041 aacttgcttc ctagtgaaac cgttcaggac atctacggga ttgttgctaa gaaagtcaac
5101 gagattctac aagcagacgc aatcaatggg accgataacg aagtagttac cgtgaccgat
5161 gagaacactg gtgaaatctc tgagaaagtc aagctgggca ctaaggcact ggctggtcaa
5221 tggctggctt acggtgttac tcgcagtgtg actaagcgtt cagtcatgac gctggcttac
5281 gggtccaaag agttcggctt ccgtcaacaa gtgctggaag ataccattca gccagctatt
5341 gattccggca agggtctgat gttcactcag ccgaatcagg ctgctggata catggctaag
5401 ctgatttggg aatctgtgag cgtgacggtg gtagctgcgg ttgaagcaat gaactggctt
5461 aagtctgctg ctaagctgct ggctgctgag gtcaaagata agaagactgg agagattctt
5521 cgcaagcgtt gcgctgtgca ttgggtaact cctgatggtt tccctgtgtg gcaggaatac
5581 aagaagccta ttcagacgcg cttgaacctg atgttcctcg gtcagttccg cttacagcct
5641 accattaaca ccaacaaaga tagcgagatt gatgcacaca aacaggagtc tggtatcgct
5701 cctaactttg tacacagcca agacggtagc caccttcgta agactgtagt gtgggcacac
5761 gagaagtacg gaatcgaatc ttttgcactg attcacgact ccttcggtac cattccggct
5821 gacgctgcga acctgttcaa agcagtgcgc gaaactatgg ttgacacata tgagtcttgt
5881 gatgtactgg ctgatttcta cgaccagttc gctgaccagt tgcacgagtc tcaattggac
5941 aaaatgccag cacttccggc taaaggtaac ttgaacctcc gtgacatctt agagtcggac
6001 ttcgcgttcg cgtaaaagct taattagctg agcttggact cctgttgata gatccagtaa
6061 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
6121 ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc taaaatggag
6181 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
6241 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
6301 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
6361 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
6421 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
6481 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
6541 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
6601 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
6661 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
6721 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
6781 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt
6841 taaggcagtt attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca
6901 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
6961 gaacgctctc ctgagtagga caaatccgcc ctctagatta cgtgcagtcg atgataagct
7021 gtcaaacatg agaattgtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
7081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
7141 gcggggagag gcgg
//
