| 质粒名称: | pQTEV-STX10 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 31572 |
| 存储实验室: | Konrad Buessow |
|
载体骨架基本信息 |
|
| 载体名称: | pQTEV |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 5513 bp |
| 载体类型: | 细菌细胞表达 |
| 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: | yntaxin 10 / PSFEp250D102 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 813bp |
| GenBank编号 | DQ000552 |
| 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(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 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-STX10 was a gift from Konrad Buessow (Addgene plasmid # 31572 ; http://n2t.net/addgene:31572 ; RRID:Addgene_31572) |
| 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. 10.1186/1475-2859-4-21 PubMed 15998469 |
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 pQTEV-STX10 5513 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pQTEV-STX10
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5513)
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 5513)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5513
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..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 970..1064
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 1108..1767
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(1174..1193)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 1832..1918
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2011..3093)
/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 3055..3074
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(3094..3171)
/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(3255..3277)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3363..3503
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 3418..3437
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(3518..3535)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3689..4277)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3769..3788)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4448..5308)
/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 5071..5090
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5309..5413)
/gene="bla"
/label=AmpR promoter
primer_bind 5481..5499
/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 cagggatcct ctctcgaaga cccctttttt gtagtccgag gcgaggtgca gaaggcggtg
241 aacacggccc gcgggctgta ccagcgctgg tgcgagctcc tgcaggaaag cgcggcggtc
301 ggacgcgagg agctggactg gacgaccaat gagctgcgga atggcctgcg cagcatcgag
361 tgggacctcg aggacctgga agagaccatc ggtatagtgg aagccaaccc aggcaagttc
421 aagctcccag ccggggacct gcaggagaga aaggtgttcg tggagcggat gcgagaggca
481 gtccaggaaa tgaaggacca tatggtcagc ccaacagccg tagcattttt ggagaggaat
541 aacagagaga tactcgcagg caagccagct gcccagaagt cacccagcga cctgctggat
601 gccagcgcag tctcggccac atctcgctac atcgaggagc agcaggccac acagcagctg
661 atcatggatg aacaggatca acagctggag atggtgtctg ggagcatcca ggttctgaag
721 cacatgtccg gccgcgttgg agaagagctg gacgagcagg gcatcatgct ggatgccttc
781 gcccaagaga tggaccacac ccagtcccgc atggacgggg tcctcaggaa gttggccaaa
841 gtatcccaca tgacgagtgg tgagtcccct caggggaggg gtcagtcctg gtgggggcag
901 gttgtaggtg gtaccctctc cccgtgagcg gccgctcgac ctgcagccaa gcttaattag
961 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
1021 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagcttggcg
1081 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
1141 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
1201 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
1261 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
1321 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
1381 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
1441 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
1501 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
1561 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
1621 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
1681 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
1741 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
1801 cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga
1861 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
1921 gccctctaga ttacgtgcag tcgatgataa gctgtcaaac atgagaattg tgcctaatga
1981 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
2041 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
2101 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
2161 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
2221 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
2281 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
2341 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
2401 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
2461 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
2521 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
2581 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
2641 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
2701 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
2761 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
2821 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
2881 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
2941 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
3001 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
3061 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
3121 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt cggaatttcg
3181 ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga gctgcctcgc gcgtttcggt
3241 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
3301 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
3361 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
3421 catcagagca gattgtactg agagtgcacc acatgcggtg tgaaataccg cacagatgcg
3481 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
3541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
3601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
3661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
3721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
3781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
3841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
3901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
3961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
4021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
4081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
4141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
4201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
4261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
4321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
4381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
4441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
4501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
4561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
4621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
4681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
4741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
4801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
4861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
4921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
4981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
5041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
5101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
5161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
5221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
5281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
5341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
5401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
5461 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//
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