| 质粒名称: | Hs.KRAS4b G12D |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 83131 |
| 存储实验室: | Dominic Esposito |
|
载体骨架基本信息 |
|
| 载体名称: | pDonor-255 |
| 载体出品公司: | -- |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 2693 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | Gateway Entry Clone |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | KRAS |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 567 bp |
| 基因突变: | G12D |
| GenBank编号 | NM_004985.4 |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13F 23-mer (5'-CCCAGTCACGACGTTGTAAAACG) |
| 3' 测序引物 | attLR 23-mer (5'-TAACATCAGAGATTTTGAGACAC) |
|
文献引用方法 |
|
| 材料和方法部分: | Hs.KRAS4b G12D was a gift from Dominic Esposito (Addgene plasmid # 83131 ; http://n2t.net/addgene:83131 ; RRID:Addgene_83131) |
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 Hs.KRAS4b G12D 3260 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Gateway ORF Entry clone of human KRAS4B [NM_004985.4 ] with stop
codon (for native or N-terminal fusions), G12D mutation.
ACCESSION .
VERSION .
KEYWORDS Hs.KRAS4b G12D
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3260)
AUTHORS Esposito
TITLE RAS mutant collection V2.0
JOURNAL unpublished
REFERENCE 2 (bases 1 to 3260)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3260
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 23..45
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 37..54
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 38..54
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 84..183
/gene="mutant version of attL"
/label=attL1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 186..752
/codon_start=1
/product="human oncoprotein generated by the G12V mutation
in the small GTPase K-Ras"
/label=K-Ras (G12V)
/translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
RKHKEKMSKDGKKKKKKSKTKCVIM"
CDS 186..752
/codon_start=1
/product="human small GTPase proto-oncoprotein"
/label=K-Ras
/translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
RKHKEKMSKDGKKKKKKSKTKCVIM"
protein_bind complement(751..850)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 1227..2018
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 2111..2699
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2600..2619
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2853..2870
/label=L4440
/note="L4440 vector, forward primer"
terminator 3029..3056
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 3148..3234
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
181 gcaccatgac tgaatataaa cttgtggtag ttggagctga tggcgtaggc aagagtgcct
241 tgacgataca gctaattcag aatcattttg tggacgaata tgatccaaca atagaggatt
301 cctacaggaa gcaagtagta attgatggag aaacctgtct cttggatatt ctcgacacag
361 caggtcaaga ggagtacagt gcaatgaggg accagtacat gaggactggg gagggctttc
421 tttgtgtatt tgccataaat aatactaaat catttgaaga tattcaccat tatagagaac
481 aaattaaaag agttaaggac tctgaagatg tacctatggt cctagtagga aataaatgtg
541 atttgccttc tagaacagta gacacaaaac aggctcagga cttagcaaga agttatggaa
601 ttccttttat tgaaacatca gcaaagacaa gacagggtgt tgatgatgcc ttctatacat
661 tagttcgaga aattcgaaaa cataaagaaa agatgagcaa agatggtaaa aagaagaaaa
721 agaagtcaaa gacaaagtgt gtaattatgt aatcaacttt cttgtacaaa gttggcatta
781 taggaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
841 tcattatttg catgcaagct tagatcctct agaccagcca ggacagaaat gcctcgactt
901 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
961 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
1021 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
1081 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
1141 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
1201 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
1261 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
1321 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
1381 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
1441 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
1501 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
1561 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
1621 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
1681 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
1741 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
1801 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
1861 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
1921 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
1981 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
2041 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
2101 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
2161 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
2221 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
2281 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
2341 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
2401 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
2461 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
2521 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
2581 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
2641 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
2701 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
2761 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
2821 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
2881 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
2941 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
3001 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
3061 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
3121 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
3181 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
3241 ggcagttccc tactctcgcg
//
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