| 质粒名称: | hPRL2_1-163 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 209794 |
| 存储实验室: | Kalle Gehring |
|
载体骨架基本信息 |
|
| 载体名称: | pET15b |
| 载体出品公司: | Novagen |
| 空载体大小: | 5708bp |
| 完整质粒大小: | 6198bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PRL-2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 951 bp |
| 基因突变: |
aa 1-163 only
Entrez GenePTP4A2 (a.k.a. HH13, HH7-2, HU-PP-1, OV-1, PRL-2, PRL2, PTP4A, PTPCAAX2, ptp-IV1a, ptp-IV1b)
|
| 启动子: | T7 |
| 标签/融合蛋白: | 6xHis (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NdeI (not destroyed) |
| 3’克隆位点: | NBamHI (not destroyed) |
| 5’测序引物 | T7 Fwd |
| 3’测序引物 | T7 Rev |
|
文献引用方法 |
|
| 材料和方法部分: | hPRL2_1-163 was a gift from Kalle Gehring (Addgene plasmid # 209794 ; http://n2t.net/addgene:209794 ; RRID:Addgene_209794) |
| 参考文献部分: | PRL3 pseudophosphatase activity is necessary and sufficient to promote metastatic growth. Kozlov G, Funato Y, Chen YS, Zhang Z, Illes K, Miki H, Gehring K. J Biol Chem. 2020 Aug 14;295(33):11682-11692. doi: 10.1074/jbc.RA120.014464. Epub 2020 Jun 22. 10.1074/jbc.RA120.014464 PubMed 32571875 |
买家导航
hPRL2_1-163载体质粒基本信息
hPRL2_1-163质粒图谱载体图谱和hPRL2_1-163载体序列质粒序列多克隆位点信息
hPRL2_1-163质粒载体简介
hPRL2_1-163质粒序列载体序列
LOCUS hPRL2_1-163 6198 bp DNA circular SYN 27-AUG-2024
DEFINITION Bacterial expression of human PRL-2 without farnesylation motif.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6198)
AUTHORS Kozlov G, Funato Y, Chen YS, Zhang Z, Illes K, Miki H, Gehring K
TITLE PRL3 pseudophosphatase activity is necessary and sufficient to
promote metastatic growth.
JOURNAL J Biol Chem. 2020 Aug 14;295(33):11682-11692. doi:
10.1074/jbc.RA120.014464. Epub 2020 Jun 22.
PUBMED 32571875
REFERENCE 2 (bases 1 to 6198)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 27, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1074/jbc.RA120.014464"; journalName: "J Biol Chem"; date:
"2020-08-14- 14"; volume: "295"; issue: "33"; pages: "11682-11692"
FEATURES Location/Qualifiers
source 1..6198
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 10..38
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
terminator complement(213..260)
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 256..274
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(829..846)
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS complement(856..873)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 892..914
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 929..953
/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)."
primer_bind complement(953..972)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(954..972)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1056..1075)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 1281..1358
/gene="lacI"
/label=lacI promoter
CDS 1359..2441
/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 complement(1378..1397)
/label=LacI-R
/note="LacI, reverse primer"
CDS 3753..3944
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(3938..3960)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 4046..4185
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4101..4120
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4200..4217)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4371..4959)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4451..4470)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5130..5990)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 5753..5772
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5991..6095)
/gene="bla"
/label=AmpR promoter
primer_bind 6163..6181
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
301 cgggctttgt tagcagccgg atcctcaatg cccattggta tctctgaagc gtaatcgcat
361 cttaggtcgg tatttctcca aataaagcag ctgtttggaa ttgaacgctc cccttctttt
421 ttgtcttata aactgaactg catcttcgta cttcattcca cattcaatca aagcaagtgc
481 aaccagcaca ggtgcccttc ccaatcctgc aacacaatgc actgcaacac agcaacctgg
541 ctcttcacga aatttggttt ttaacaggtt taaccaatca tctactatct gattaggggg
601 tggagctcca tcatcaaatg gccaatctag aacgtggatt ccttcttttt caactggagc
661 tttatcatat gtagcatcac aaactcgaac caaagtcgtc actccatact tcttaagttc
721 ctctgtgaac ttgttgagag tagcattggt agggttgtga gttatcagaa aacgcatgtt
781 ctcataggag atctccacag gggctggacg gttcatctcg agcatatggc tgccgcgcgg
841 caccaggccg ctgctgtgat gatgatgatg atggctgctg cccatggtat atctccttct
901 taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag
961 tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc
1021 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
1081 ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc
1141 aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc
1201 ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa
1261 gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
1321 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
1381 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
1441 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
1501 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
1561 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
1621 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
1681 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
1741 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
1801 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
1861 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
1921 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
1981 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
2041 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
2101 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
2161 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
2221 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
2281 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
2341 ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
2401 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
2461 agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca
2521 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
2581 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
2641 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
2701 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
2761 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
2821 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
2881 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
2941 ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac ggcgatttat
3001 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt
3061 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa
3121 gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc
3181 ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct
3241 ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga
3301 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
3361 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
3421 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
3481 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
3541 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
3601 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
3661 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
3721 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
3781 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
3841 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
3901 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
3961 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
4021 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
4081 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg
4141 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
4201 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
4261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
4321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
4381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
4441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
4501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
4561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
4621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
4681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
4741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
4801 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
4861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
4921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
4981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
5041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
5101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
5161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
5221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
5281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
5341 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
5401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
5461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
5521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
5581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
5641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
5701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
5761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
5821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
5881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
5941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
6001 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
6061 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
6121 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
6181 ccctttcgtc ttcaagaa
//
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