| 质粒名称: | pBabe-Kras G12D |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 58902 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pBabe-puro |
| 载体出品公司: | 未知 |
| 空载体大小: | 5169bp |
| 完整质粒大小: | 5683 bp |
| 载体类型: | 哺乳细胞表达、Retroviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | KRAS /K-Ras2A / RASK2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 600 bp |
| 基因突变: | G12D mutation |
| GenBank编号 | NM_004985.4 NP_004976.2 |
| 标签/融合蛋白: | HA (N terminal on insert) |
|
文献引用方法 |
|
| 材料和方法部分: | pBabe-Kras G12D was a gift from Channing Der (Addgene plasmid # 58902 ; http://n2t.net/addgene:58902 ; RRID:Addgene_58902) |
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 pBabe-Kras G12D 5683 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses human Kras with a G12D Mutation.
ACCESSION .
VERSION .
KEYWORDS pBabe-Kras G12D
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5683)
TITLE Mutant Ras Constructs
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5683)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5683
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 120..589
/note="long terminal repeat from Moloney murine leukemia
virus"
CDS complement(810..1409)
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(893..913)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 1390..1409
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
promoter complement(1419..1748)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1433..1568
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(1487..1506)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 1733..1753
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
CDS complement(1765..2331)
/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 complement(1765..2331)
/codon_start=1
/product="human small GTPase proto-oncoprotein"
/label=K-Ras
/translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
RKHKEKMSKDGKKKKKKSKTKCVIM"
CDS complement(2359..2385)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(2408..2824)
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind complement(2424..2440)
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(2456..2478)
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
misc_feature 3025..3224
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
LTR 3289..3758
/note="long terminal repeat from Moloney murine leukemia
virus"
promoter 3789..3893
/gene="bla"
/label=AmpR promoter
CDS 3894..4754
/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(4112..4131)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4914..5502
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5403..5422
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5656..5673
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg
61 gccatttaat gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt
121 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
181 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact
241 cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg
301 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
361 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
421 tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
481 cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag
541 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct
601 gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa
661 aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt
721 ttctggagac taaataaaat cttttatttt atcgatgcat ggggtcgtgc gctcctttcg
781 gtcgggcgct gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag
841 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
901 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
961 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
1021 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
1081 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
1141 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
1201 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
1261 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
1321 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
1381 aggcgcaccg tgggcttgta ctcggtcatg gtaagctttt tgcaaaagcc taggcctcca
1441 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
1501 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
1561 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
1621 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
1681 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
1741 tgacacacat tccacagggt cgacttacat aattacacac tttgtctttg acttcttttt
1801 cttcttttta ccatctttgc tcatcttttc tttatgtttt cgaatttctc gaactaatgt
1861 atagaaggca tcatcaacac cctgtcttgt ctttgctgat gtttcaataa aaggaattcc
1921 ataacttctt gctaagtcct gagcctgttt tgtgtctact gttctggaag gcaaatcaca
1981 tttatttcct actaggacca taggtacatc ttcagagtcc ttaactcttt taatttgttc
2041 tctataatgg tgaatatctt caaatgattt agtattattt atggcaaata cacaaagaaa
2101 gccctcccca gtcctcatgt actggtccct cattgcactg tactcctctt gacctgctgt
2161 gtcgagaata tccaagagac aggtttctcc atcaattact acttgcttcc tgtaggaatc
2221 ctctattgtt ggatcatatt cgtccacaaa atgattctga attagctgta tcgtcaaggc
2281 actcttgcct acgccgtcag ctccaactac cacaagttta tattcagtca tggtggatcc
2341 agagggtcct ccaagtgagg catagtcggg cacatcatat ggataacttg aagccattga
2401 tccggccggc gcctagagaa ggagtgaggg ctggataaag ggaggatcga ggcggggtcg
2461 aacgaggagg ttcaaggggg agagacgggg cggatggagg aagaggaggc ggaggcttag
2521 ggtgtacaaa gggcttgacc cagggagggg ggtcaaaagc caaggcttcc caggtcacga
2581 tgtaggggac ctggtctggg tgtccatgcg ggccaggtga aaagaccttg atcttaacct
2641 gggtgatgag gtctcggtta aaggtgccgt ctcgcggcca tccgacgtta aaggttggcc
2701 attctgcaga gcagaaggta acccaacgtc tcttcttgac atctaccgac tggttgtgag
2761 cgatccgctc gacatctttc cagtgaccta aggtcaaact taagggagtg gtaacagtct
2821 ggccctaatt ttcagacaaa tacagaaaca cagtcagaca gagacaacac agaacgatgc
2881 tgcagcgctg cagcagacaa gacgcgcggc ttcggttcca aaccgaaagc aaaaattcag
2941 acggaggcgg gaactgtttt aggttctcgt ctcctaccag aaccacatat cctgacgggg
3001 tcggattcca catcgactcc cttcctcagg tcgggccaca aaaacggccc ccaaagtccc
3061 tgggacgtct cccagggttg cggccgggtg ttcagaactc gtcagttcca ccacgggtcc
3121 gccagataca gagctagtta gctaactagt accgacgcag gcgcataaaa tcagtcatag
3181 acactagaca atcggacaga cacagataag ttgctggcca gcttgcctcc cggtggtggg
3241 tcggtggtcc ctgggcaggg gtctcccgat cccggacgag cccccaaatg aaagaccccc
3301 gctgacgggt agtcaatcac tcagaggaga ccctcccaag gaacagcgag accacaagtc
3361 ggatgcaact gcaagagggt ttattggata cacgggtacc cgggcgactc agtcaatcgg
3421 aggactggcg ccccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc
3481 gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt
3541 caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct gagggctgga
3601 ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga
3661 tatcctgttt ggcccatatt cagctgttcc atctgttctt ggccctgagc cggggcagga
3721 actgcttacc acagatatcc tgtttggccc atattcaggc tgcaggtggc acttttcggg
3781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
3841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
3901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
3961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
4021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
4081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
4141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
4201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
4261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
4321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
4381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
4441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
4501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
4561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
4621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
4681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
4741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttgcggc
4801 cgcaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
4861 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
4921 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
4981 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
5041 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
5101 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
5161 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
5221 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
5281 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
5341 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
5401 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
5461 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
5521 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
5581 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
5641 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
