| 質粒名稱: | pBSFI-myr-akt-T308A/S473A |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 49191 |
| 存儲實驗室: | Peter Vogt |
|
載體骨架基本信息 |
|
| 載體名稱: | pBSFI |
| 載體出品公司: | -- |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | 3000 bp |
| 完整質粒大小: | 4500 bp |
| 載體修飾: | pBSFI基于pBluescript SK，在SacI和KpnI之間插入了不同的MCS |
| 載體類型: | Adaptor Plasmid |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | Akt1,Akt |
| 物種來源: | M. musculus (mouse) |
| 插入大小: | 1500 bp |
| 基因突變: | T308A; S473A |
| GenBank編號 | NM_009652.3 |
| 啟動子 | T7 |
| 標簽/融合蛋白: |
myristolation signal（插入件上的N端子）
HA tag （插接件上的C端子）
|
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | BamHI（克隆過程中銷毀） |
| 3' 克隆位點: | EcoRI（未銷毀） |
| 5' 測序引物 | T3 |
| 3' 測序引物 | T7 |
|
文獻引用方法 |
|
| 材料和方法部分: | pBSFI-myr-akt-T308A/S473A was a gift from Peter Vogt (Addgene plasmid # 49191 ; //n2t.net/addgene:49191 ; RRID:Addgene_49191) |
| 參考文獻部分: | The akt kinase: molecular determinants of oncogenicity. Aoki M, Batista O, Bellacosa A, Tsichlis P, Vogt PK. Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14950-5. 10.1073/pnas.95.25.14950 PubMed 9843996 |
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pBSFI-myr-akt-T308A/S473A載體質粒基本信息
pBSFI-myr-akt-T308A/S473A質粒圖譜載體圖譜和pBSFI-myr-akt-T308A/S473A載體序列質粒序列多克隆位點信息
pBSFI-myr-akt-T308A/S473A質粒載體簡介
pBSFI-myr-akt-T308A/S473A質粒序列載體序列
LOCUS pBSFI-myr-akt-T308A/S473A 4474 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cDNA of Akt1 with myristyolation signal and point mutation from T to
A at position 308 and S to A at position 473 in cloning vector.
ACCESSION .
VERSION .
KEYWORDS pBSFI-myr-akt-T308A/S473A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4474)
AUTHORS Aoki M, Batista O, Bellacosa A, Tsichlis P, Vogt PK
TITLE The akt kinase: molecular determinants of oncogenicity.
JOURNAL Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14950-5.
PUBMED 9843996
REFERENCE 2 (bases 1 to 4474)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..4474
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1536..1562
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(1631..1650)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1632..1650)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1657..1674)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1657..1673)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1666..1688)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(1814..2269)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1951..1972)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2163..2182
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2296..2400
/gene="bla"
/label=AmpR promoter
CDS 2401..3261
/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 complement(2619..2638)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3432..4020
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3921..3940
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4174..4191
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4308..4329
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4344..4374
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4382..4398
/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 4387..4409
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4406..4422
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4406..4422
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 4443..4461
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 agctggccat tacggccgcg gccgcctcta gaggatctgg taccacgcgt atcgataagc
61 ttatggggag cagcaagagc aagcccaagt ctagaatgaa cgacgtagcc attgtgaagg
121 agggctggct gcacaaacga ggggaatata ttaaaacctg gcggccacgc tacttcctcc
181 tcaagaacga tggcaccttt attggctaca aggaacggcc tcaggatgtg gatcagcgag
241 agtccccact caacaacttc tcagtggcac aatgccagct gatgaagaca gagcggccaa
301 ggcccaacac ctttatcatc cgctgcctgc agtggaccac agtcattgag cgcaccttcc
361 atgtggaaac gcctgaggag cgggaagaat gggccaccgc cattcagact gtggccgatg
421 gactcaagag gcaggaagaa gagacgatgg acttccgatc aggctcaccc agtgacaact
481 caggggctga agagatggag gtgtccctgg ccaagcccaa gcaccgtgtg accatgaacg
541 agtttgagta cctgaaacta ctgggcaagg gcacctttgg gaaagtgatt ctggtgaaag
601 agaaggccac aggccgctac tatgccatga agatcctcaa gaaggaggtc atcgtcgcca
661 aggatgaggt tgcccacacg cttactgaga accgtgtcct gcagaactct aggcatccct
721 tccttacggc cctcaagtac tcattccaga cccacgaccg cctctgcttt gtcatggagt
781 atgccaacgg gggcgagctc ttcttccacc tgtctcgaga gcgcgtgttc tccgaggacc
841 gggcccgctt ctatggtgcg gagattgtgt ctgccctgga ctacttgcac tccgagaaga
901 acgtggtgta ccgggacctg aagctggaga acctcatgct ggacaaggac gggcacatca
961 agataacgga cttcgggctg tgcaaggagg ggatcaagga tggtgccact atgaaggcat
1021 tctgcggaac gccggagtac ctggcccctg aggtgctgga ggacaacgac tacggccgtg
1081 cagtggactg gtgggggctg ggcgtggtca tgtatgagat gatgtgtggc cgcctgccct
1141 tctacaacca ggaccacgag aagctgttcg agctgatcct catggaggag atccgcttcc
1201 cgcgcacact cggccctgag gccaagtccc tgctctccgg gctgctcaag aaggacccta
1261 cacagaggct cggtgggggc tctgaggatg ccaaggagat catgcagcac cggttctttg
1321 ccaacatcgt gtggcaggat gtgtatgaga agaagctgag cccacctttc aagccccagg
1381 tcacctctga gactgacacc aggtatttcg atgaggagtt cacagctcag atgatcacca
1441 tcacgccgcc tgatcaagat gacagcatgg agtgtgtgga cagtgagcgg aggccgcact
1501 tcccccagtt cgcctactca gccagtggca cagcctaccc atacgatgtt ccagattacg
1561 cttctagatg aattcacgcg taagcttatc gatgtcgacg ggcccggccg cctcggccgt
1621 acccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
1681 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
1741 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
1801 gaatggcgaa tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
1861 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
1921 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
1981 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
2041 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
2101 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
2161 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
2221 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
2281 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
2341 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
2401 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
2461 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
2521 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
2581 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
2641 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
2701 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
2761 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
2821 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
2881 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
2941 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
3001 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
3061 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
3121 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
3181 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
3241 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
3301 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
3361 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
3421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
3481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
3541 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
3601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
3661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
3721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
3781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
3841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
3901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
3961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
4021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
4081 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
4141 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
4201 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
4261 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
4321 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
4381 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
4441 gaaattaacc ctcactaaag ggaacaaaag ctgg
//