| 質粒名稱: | pBSFI-myr-akt1 |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 49186 |
| 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: | -- |
| 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-akt1 was a gift from Peter Vogt (Addgene plasmid # 49186 ; //n2t.net/addgene:49186 ; RRID:Addgene_49186) |
| 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-akt1載體質粒基本信息
pBSFI-myr-akt1質粒圖譜載體圖譜和pBSFI-myr-akt1載體序列質粒序列多克隆位點信息
pBSFI-myr-akt1質粒載體簡介
pBSFI-myr-akt1質粒序列載體序列
LOCUS pBSFI-myr-akt1 4477 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cDNA of Akt1 with myristyolation signal in cloning vector.
ACCESSION .
VERSION .
KEYWORDS pBSFI-myr-akt1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4477)
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 4477)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..4477
/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(1660..1677)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1660..1676)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1669..1691)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(1817..2272)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1954..1975)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2166..2185
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2299..2403
/gene="bla"
/label=AmpR promoter
CDS 2404..3264
/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(2622..2641)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3435..4023
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3924..3943
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4177..4194
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4311..4332
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4347..4377
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4385..4401
/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 4390..4412
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4409..4425
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4409..4425
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 4446..4464
/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 atgaagacat
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 ctcctactca gccagtggca cagcctaccc atacgatgtt ccagattacg
1561 cttctagatg aattcacgcg taagcttatc gatgtcgacg ggcccggccg cctcggccgt
1621 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
1681 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
1741 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
1801 cctgaatggc gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
1861 tgttaaatca gctcattttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
1921 aaaagataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
1981 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
2041 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
2101 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
2161 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
2221 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
2281 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
2341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
2401 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
2461 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
2521 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
2581 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
2641 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
2701 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
2761 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
2821 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
2881 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
2941 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
3001 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
3061 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
3121 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
3181 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
3241 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
3301 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
3361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
3421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
3481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
3541 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
3601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
3661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
3721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
3781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
3841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
3901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
3961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
4021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
4081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
4141 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
4201 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
4261 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
4321 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
4381 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
4441 ctcgaaatta accctcacta aagggaacaa aagctgg
//
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