| 质粒名称: | pMKO.1 GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 10676 |
| 存储实验室: | William Hahn |
|
载体骨架基本信息 |
|
| 载体名称: | pMKO.1 |
| 载体出品公司: | -- |
| 空载体大小: | 6700 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,逆转录病毒，RNAi |
| 筛选标记: | GFP |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | pLXSN 5' |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pMKO.1 GFP was a gift from William Hahn (Addgene plasmid # 10676 ; http://n2t.net/addgene:10676 ; RRID:Addgene_10676) |
买家导航
pMKO.1GFP载体基本信息
pMKO.1GFP载体图谱质粒图谱和多克隆位点信息
pMKO.1GFP载体简介
pMKO.1GFP载体序列
LOCUS pMKO.1 GFP 6767 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMKO.1 GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6767)
TITLE Viral vectors.
REFERENCE 2 (bases 1 to 6767)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6767
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(197..216)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter complement(238..567)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 252..387
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 708..730
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
LTR 833..1258
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
CDS complement(1410..2129)
/codon_start=1
/product="humanized Renilla green fluorescent protein"
/label=hrGFP
/note="mammalian codon-optimized"
/translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
AQLTSLGKPLGSLHEWV"
primer_bind 2134..2150
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 2135..2151
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2160..2489)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2174..2309
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(2558..2798)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(2608..2627)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(2778..2798)
/label=hU6-F
/note="Human U6 promoter, forward primer"
CDS complement(2834..3250)
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind complement(2850..2866)
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(2882..2904)
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
misc_feature 3451..3650
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
LTR 3713..3888
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
promoter complement(3889..4092)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3918..3938)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 4093..4472
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind complement(4572..4590)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4868..4972
/gene="bla"
/label=AmpR promoter
CDS 4973..5833
/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(5191..5210)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6004..6592
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6493..6512
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ccatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
61 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
121 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
181 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag taagcttttt
241 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
301 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
361 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
421 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
481 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
541 ggactttcca caccctaact gacacacatt ccacagccgg atctacactc cgctatcgct
601 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
661 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
721 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagccagctt acctcccggt
781 ggtgggtcgg tggtccctgg gcaggggtct cccgatcccg gacgagcccc caaatgaaag
841 acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac agcgagacca
901 caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg cgactcagtc
961 aatcggagga ctggcgcccc gagtgagggg ttgtgggctc ttttattgag ctcggggagc
1021 agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata aggcacaggg
1081 tcatttcagg tccttggggc accctggaaa catctgatgg ttctctaggg tcgaccctgt
1141 tccatctgtt cctgaccttg atctgaactt ctctattctc agttatgtat ttttccatgc
1201 cttgcaaaat ggcgttactt aagctagctt gccaaaccta caggtggggt ctttcattcc
1261 cccctttttc tggagactaa ataaaatctt ttattttatc tatggctcgt actctatagg
1321 cttcagctgg tgatattgtt gagtcaaaac tagagcctgg accactgata tcctgtcttt
1381 aacaaattgg actaatcgaa ggtaccctat tacacccact cgtgcaggct gcccaggggc
1441 ttgcccaggc tggtcagctg ggcgatggcg gtctcgtgct gctccacgaa gccgccgtcc
1501 tccacgtagg tcttctccag gcggtgctgg atgaagtggt actcggggaa gtccttcacc
1561 acgcccttgc tcttcatcag ggtgcgcatg tggcagctgt agaacttgcc gctgttcagg
1621 cggtacacca ggatcacctg gcccaccagc acgccgtcgt tcatgtacac cacctcgaag
1681 ctgggctgca ggccggtgat ggtcttcttc atcacggggc cgtcgttggg gaagttgcgg
1741 cccttgtact ccacgcggta cacgaacatc tcctcgatca ggttgatgtc gctgcggatc
1801 tccaccaggc cgccgtcctc gtagcgcagg gtgcgctcgt acacgaagcc ggcggggaag
1861 ctctggatga agaagtcgct gatgtcctcg gggtacttgg tgaaggtgcg gttgccgtac
1921 tggaaggcgg ggctcaggat gtcgaaggcg aagggcaggg gggcgccctt ggtcacgcgg
1981 atctgcacca gctggttgcc gaacaggatg ttgcccttgc cgcagccctc catggtgaac
2041 acgtggttgt tcaccacgcc ctccaggttc accttgaagc tcatgatctc ctgcaggccg
2101 gtgttcttca ggatctgctt gctcaccatg gtctcgaggt cgacggtatc gataagcttt
2161 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
2221 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
2281 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
2341 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
2401 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
2461 ggggactttc cacaccctaa ctgacacaca ttccacaggg tcgaccactg tgctggcgaa
2521 ttccatatgg gagtactagt acgcgtaccg gtgtttcgtc ctttccacaa gatatataaa
2581 gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat
2641 aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact
2701 aatatctttg tgtttgcagt caaattaatt ctaattatct ctctaacagc cttgtatcgt
2761 atatgcaaat atgaaggaat catgggaaat aggccctctt cctgcccgac cttgtcgaca
2821 gatcttcaat tccggcgcct agagaaggag tgagggctgg ataaagggag gatcgaggcg
2881 gggtcgaacg aggaggttca agggggagag acggggcgga tggaggaaga ggaggcggag
2941 gcttagggtg tacaaagggc ttgacccagg gaggggggtc aaaagccaag gcttcccagg
3001 tcacgatgta ggggacctgg tctgggtgtc catgcgggcc aggtgaaaag accttgatct
3061 taacctgggt gatgaggtct cggttaaagg tgccgtctcg cggccatccg acgttaaagg
3121 ttggccattc tgcagagcag aaggtaaccc aacgtctctt cttgacatct accgactggt
3181 tgtgagcgat ccgctcgaca tctttccagt gacctaaggt caaacttaag ggagtggtaa
3241 cagtctggcc ctaattttca gacaaataca gaaacacagt cagacagaga caacacagaa
3301 cgatgctgca gcgctgcagc agacaagacg cgcggcttcg gttccaaacc gaaagcaaaa
3361 attcagacgg aggcgggaac tgttttaggt tctcgtctcc taccagaacc acatatcctg
3421 acggggtcgg attccacatc gactcccttc ctcaggtcgg gccacaaaaa cggcccccaa
3481 agtccctggg acgtctccca gggttgcggc cgggtgttca gaactcgtca gttccaccac
3541 gggtccgcca gatacagagc tagttagcta actagtacag acgcaggcgc ataacatcaa
3601 acatagacac tagacaatcg gacagacaca gataagttgc tggccagctt acctcccggt
3661 ggtgggtcgg tggtccctgg gcaggggtct ccaaatcccg gacgagcccc caaatgaaag
3721 acccccgtcg tgggtagtca atcactcaga ggagaccctc ccaaggaaca gcgagaccac
3781 gattcggatg caaacagcaa gaggctttat tgggaatacg ggtacccggg cgacgcagtc
3841 tatcggaaga ctggcgcgcc gagtgagggg ttgtgggctc ttttattgag ctctgcttat
3901 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
3961 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
4021 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
4081 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
4141 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
4201 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
4261 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
4321 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
4381 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
4441 ttactattaa taactagtca ataatcaatg tcaacatggc ggtaatgttg gacatgagcc
4501 aatataaatg tacatattat gatatggata caacgtatgc aatggccaag ctaattcttg
4561 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
4621 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
4681 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
4741 ataattaagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
4801 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
4861 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
4921 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
4981 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
5041 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
5101 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
5161 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
5221 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
5281 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
5341 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
5401 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
5461 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
5521 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
5581 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
5641 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
5701 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
5761 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
5821 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
5881 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
5941 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
6001 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
6061 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
6121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
6181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
6241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
6301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
6361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
6421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
6481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
6541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
6601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
6661 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
6721 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaag
//
