| 质粒名称: | 800 pSG5L HA PTEN wt |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 10750 |
| 存储实验室: | William Sellers |
|
载体骨架基本信息 |
|
| 载体名称: | pSG5L-HA |
| 空载体大小: | 4100 bp |
| 完整质粒大小: | 5367 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | PTEN |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1200 bp |
| 标签/融合蛋白: | HA (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: | EcoRI(未破坏) |
| 5’测序引物 | T7 |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | 800 pSG5L HA PTEN wt was a gift from William Sellers (Addgene plasmid # 10750 ; http://n2t.net/addgene:10750 ; RRID:Addgene_10750) |
| 参考文献部分: | Regulation of G1 progression by the PTEN tumor suppressor protein is linked to inhibition of the phosphatidylinositol 3-kinase/Akt pathway. Ramaswamy S, Nakamura N, Vazquez F, Batt DB, Perera S, Roberts TM, Sellers WR. Proc Natl Acad Sci U S A. 1999 Mar 2. 96(5):2110-5. 10.1073/pnas.96.5.2110 PubMed 10051603 |
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 800 pSG5L HA PTEN wt 5367 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS 800 pSG5L HA PTEN wt
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5367)
AUTHORS Ramaswamy S, Nakamura N, Vazquez F, Batt DB, Perera S, Roberts TM,
Sellers WR
TITLE Regulation of G1 progression by the PTEN tumor suppressor protein is
linked to inhibition of the phosphatidylinositol 3-kinase/Akt
pathway.
JOURNAL Proc Natl Acad Sci U S A. 1999 Mar 2. 96(5):2110-5.
PUBMED 10051603
REFERENCE 2 (bases 1 to 5367)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5367
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(30..52)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 152..171
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(238..693)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(375..396)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 587..606
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 819..841
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 833..850
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 834..850
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(887..907)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 995..1205
/label=SV40 promoter
/note="SV40 early promoter"
promoter 1017..1213
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1064..1199
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1126..1145
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
intron 1244..1816
/label=-beta-globin intron
/note="intron from rabbit beta-globin gene"
primer_bind 1824..1843
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
primer_bind 1871..1890
/label=T7
/note="T7 promoter, forward primer"
promoter 1871..1889
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1923..1949
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 3223..3344
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3260..3279)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3314..3333
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3378..3395)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3549..4137)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3629..3648)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4308..5168)
/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 4931..4950
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5169..5273)
/gene="bla"
/label=AmpR promoter
primer_bind 5341..5359
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
241 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
301 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
361 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
421 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
481 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
541 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
601 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
661 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
721 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
781 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
841 gacggccagt gaattgtcga cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
901 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
961 aattagtcag caaggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
1021 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
1081 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
1141 cgaggccgcc tcggcctctg agctattcca gaagtagtga agaggctttt ttggaggcct
1201 aggcttttgc aaaaagctcc ggatcgatcc tgagaacttc agggtgagtt tggggaccct
1261 tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg gcaaagtttt
1321 cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catggaccct catgataatt
1381 ttgtttcttt cactttctac tctgttgaca accattgtct cctcttattt tcttttcatt
1441 ttctgtaact ttttcgttaa actttagctt gcatttgtaa cgaattttta aattcacttt
1501 tgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag gcaatcaggg
1561 tatattatat tgtacttcag cacagtttta gagaacaatt gttataatta aatgataagg
1621 tagaatattt ctgcatataa attctggctg gcgtggaaat attcttattg gtagaaacaa
1681 ctacatcctg gtcatcatcc tgcctttctc tttatggtta caatgatata cactgtttga
1741 gatgaggata aaatactctg agtccaaacc gggcccctct gctaaccatg ttcatgcctt
1801 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
1861 caaagaattg taatacgact cactataggg cgaattgtcg acccgggaag cttaccatgg
1921 cctaccccta cgacgtgccc gactacgcct ccctcggatc catgacagcc atcatcaaag
1981 agatcgttag cagaaacaaa aggagatatc aagaggatgg attcgactta gacttgacct
2041 atatttatcc aaacattatt gctatgggat ttcctgcaga aagacttgaa ggcgtataca
2101 ggaacaatat tgatgatgta gtaaggtttt tggattcaaa gcataaaaac cattacaaga
2161 tatacaatct ttgtgctgaa agacattatg acaccgccaa atttaattgc agagttgcac
2221 aatatccttt tgaagaccat aacccaccac agctagaact tatcaaaccc ttttgtgaag
2281 atcttgacca atggctaagt gaagatgaca atcatgttgc agcaattcac tgtaaagctg
2341 gaaagggacg aactggtgta atgatatgtg catatttatt acatcggggc aaatttttaa
2401 aggcacaaga ggccctagat ttctatgggg aagtaaggac cagagacaaa aagggagtaa
2461 ctattcccag tcagaggcgc tatgtgtatt attatagcta cctgttaaag aatcatctgg
2521 attatagacc agtggcactg ttgtttcaca agatgatgtt tgaaactatt ccaatgttca
2581 gtggcggaac ttgcaatcct cagtttgtgg tctgccagct aaaggtgaag atatattcct
2641 ccaattcagg acccacacga cgggaagaca agttcatgta ctttgagttc cctcagccgt
2701 tacctgtgtg tggtgatatc aaagtagagt tcttccacaa acagaacaag atgctaaaaa
2761 aggacaaaat gtttcacttt tgggtaaata cattcttcat accaggacca gaggaaacct
2821 cagaaaaagt agaaaatgga agtctatgtg atcaagaaat cgatagcatt tgcagtatag
2881 agcgtgcaga taatgacaag gaatatctag tacttacttt aacaaaaaat gatcttgaca
2941 aagcaaataa agacaaagcc aaccgatact tttctccaaa ttttaaggtg aagctgtact
3001 tcacaaaaac agtagaggag ccgtcaaatc cagaggctag cagttcaact tctgtaacac
3061 cagatgttag tgacaatgaa cctgatcatt atagatattc tgacaccact gactctgatc
3121 cagagaatga accttttgat gaagatcagc atacacaaat tacaaaagtc tgagaattcc
3181 ccgatatcgc ctcgagtcta gatccagatc ttattaaagc agaacttgtt tattgcagct
3241 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
3301 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggtcgact
3361 ctagactctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
3421 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
3481 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
3541 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
3601 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
3661 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
3721 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
3781 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
3841 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
3901 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
3961 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
4021 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
4081 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
4141 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
4201 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
4261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
4321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
4381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
4441 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
4501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
4561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
4621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
4681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
4741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
4801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
4861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
4921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
4981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
5041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
5101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
5161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
5221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
5281 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
5341 aataggcgta tcacgaggcc ctttcgt
//
