| 质粒名称: | pQTEV-YWHAE |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 31562 |
| 存储实验室: | Konrad Buessow |
|
载体骨架基本信息 |
|
| 载体名称: | pQTEV |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700bp |
| 完整质粒大小: | 5540bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Rosetta |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | PSFEp250H041、tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, epsilon、 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 840bp |
| GenBank编号 | DQ000541 |
| 启动子: | CMV |
| 标签/融合蛋白: |
His (N terminal on backbone)
TEV (N terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI (not destroyed) |
| 3’克隆位点: | NotI (not destroyed) |
| 5’测序引物 | TGAGCGGATAACAATTTCACACAG |
| 3’测序引物 | GGCAACCGAGCGTTCTGAAC |
|
文献引用方法 |
|
| 材料和方法部分: | pQTEV-YWHAE was a gift from Konrad Buessow (Addgene plasmid # 31562 ; http://n2t.net/addgene:31562 ; RRID:Addgene_31562) |
| 参考文献部分: | Structural genomics of human proteins--target selection and generation of a public catalogue of expression clones. Bussow K, Scheich C, Sievert V, Harttig U, Schultz J, Simon B, Bork P, Lehrach H, Heinemann U. Microb Cell Fact. 2005 Jul 5;4:21. doi: 10.1186/1475-2859-4-21. 10.1186/1475-2859-4-21 PubMed 15998469 |
买家导航
pQTEV-YWHAE载体质粒基本信息
pQTEV-YWHAE质粒图谱载体图谱和pQTEV-YWHAE载体序列质粒序列多克隆位点信息
pQTEV-YWHAE质粒载体简介
pQTEV-YWHAE质粒序列载体序列
LOCUS pQTEV-YWHAE 5540 bp DNA circular SYN 14-APR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5540)
AUTHORS Bussow K, Scheich C, Sievert V, Harttig U, Schultz J, Simon B, Bork
P, Lehrach H, Heinemann U
TITLE Structural genomics of human proteins--target selection and
generation of a public catalogue of expression clones.
JOURNAL Microb Cell Fact. 2005 Jul 5. 4():21.
PUBMED 15998469
REFERENCE 2 (bases 1 to 5540)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Microb Cell
Fact. 2005 Jul 5. 4():21."
COMMENT Sequence Label: pQTEV-YWHAE
FEATURES Location/Qualifiers
source 1..5540
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter join(10..24,25..30,31..47,48..54)
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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 62..78
/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 97..108
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 121..141
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
CDS 166..186
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
terminator 997..1091
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 1135..1794
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(1201..1220)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 1859..1945
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2038..3120)
/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"
primer_bind 3082..3101
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(3121..3198)
/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(3282..3304)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3390..3530
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 3445..3464
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(3545..3562)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3716..4304)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3796..3815)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(4475..5266,5267..5335))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5098..5117
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5336..5440)
/gene="bla"
/label=AmpR promoter
primer_bind 5508..5526
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaaa
121 catcaccatc accatcacca tagcgattac gacatcccca ctactgagaa tctttatttt
181 cagggatccg atgatcgaga ggatctggtg taccaggcga agctggccga gcaggctgag
241 cgatacgacg aaatggtggg gtcaatgaag aaagtagcag ggatggatgt ggagctgaca
301 gttgaagaaa gaaacctcct atctgttgca tataagaatg tgattggagc tagaagagcc
361 tcctggagaa taatcagcag cattgaacag aaagaagaaa acaagggagg agaagacaag
421 ctaaaaatga ttcgggaata tcggcaaatg gttgagactg agctaaagtt aatctgttgt
481 gacattctgg atgtactgga caaacacctc attccagcag ctaacactgg cgagtccaag
541 gttttctatt ataaaatgaa aggggactac cacaggtatc tggcagaatt tgccacagga
601 aacgacagga aggaggctgc ggagaacagc ctagtggctt ataaagctgc tagtgatatt
661 gcaatgacag aacttccacc aacgcatcct attcgcttag gtcttgctct caatttttcc
721 gtattctact acgaaattct taattcccct gaccgtgcct gcaggttggc aaaagcagct
781 tttgatgatg caattgcaga actggatacg ctgagtgaag aaagctataa ggactctaca
841 cttatcatgc agttgttacg tgataatctg acactatgga cttcagacat gcagggtgac
901 ggtgaagagc agaataaaga agcgctgcag gacgtggaag acgaaaatca gtgagcggcc
961 gctcgacctg cagccaagct taattagctg agcttggact cctgttgata gatccagtaa
1021 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
1081 ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc taaaatggag
1141 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
1201 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
1261 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
1321 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
1381 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
1441 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
1501 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
1561 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
1621 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
1681 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
1741 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt
1801 taaggcagtt attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca
1861 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
1921 gaacgctctc ctgagtagga caaatccgcc ctctagatta cgtgcagtcg atgataagct
1981 gtcaaacatg agaattgtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
2041 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
2101 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
2161 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
2221 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
2281 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
2341 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
2401 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
2461 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
2521 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
2581 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
2641 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
2701 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
2761 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
2821 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
2881 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
2941 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
3001 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
3061 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
3121 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
3181 gcaccattcg atggtgtcgg aatttcgggc agcgttgggt cctggccacg ggtgcgcatg
3241 atctagagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
3301 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
3361 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
3421 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccaca
3481 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
3541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
3601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
3661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
3721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
3781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
3841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
3901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
3961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
4021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
4081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
4141 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
4201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
4261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
4321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
4381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
4441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
4501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
4561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
4621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
4681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
4741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
4801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
4861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
4921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
4981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
5041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
5101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
5161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
5221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
5281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
5341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
5401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
5461 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
5521 cgaggccctt tcgtcttcac
//
