| 质粒名称: | pSUPER-retro-puro-shNup133-2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 87333 |
| 存储实验室: | Lei Lu |
|
载体骨架基本信息 |
|
| 载体名称: | pSUPER.retro.puro |
| 载体出品公司: | OligoEngine |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6349 bp |
| 载体类型: | 哺乳细胞表达,RNAi |
| 筛选标记: | 嘌呤霉素 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | shRNA against human Nup133 |
| 物种来源: | H. sapiens (human) |
| 启动子 | H1 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Bglll（在克隆过程中被销毁） |
| 3' 克隆位点: | XhoI (未破坏) |
| 5' 测序引物 | GGAAGCCTTGGCTTTTG |
|
文献引用方法 |
|
| 材料和方法部分: | pSUPER-retro-puro-shNup133-2 was a gift from Lei Lu (Addgene plasmid # 87333 ; http://n2t.net/addgene:87333 ; RRID:Addgene_87333) |
| 参考文献部分: | The development of a single molecule fluorescence standard and its application in estimating the stoichiometry of the nuclear pore complex. Tie HC, Madugula V, Lu L. Biochem Biophys Res Commun. 2016 Sep 30;478(4):1694-9. doi: 10.1016/j.bbrc.2016.09.005. Epub 2016 Sep 6. 10.1016/j.bbrc.2016.09.005 PubMed 27613095 |
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 pSUPER-retro-puro-shNup133-2 6351 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION To express shRNA against human Nup133.
ACCESSION .
VERSION .
KEYWORDS pSUPER-retro-puro-shNup133-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6351)
AUTHORS Tie HC, Madugula V, Lu L
TITLE The development of a single molecule fluorescence standard and its
application in estimating the stoichiometry of the nuclear pore
complex.
JOURNAL Biochem Biophys Res Commun. 2016 Sep 30;478(4):1694-9. doi:
10.1016/j.bbrc.2016.09.005. Epub 2016 Sep 6.
PUBMED 27613095
REFERENCE 2 (bases 1 to 6351)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6351
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..517
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 581..922
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 989..1405
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1335..1357
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1373..1389
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
promoter complement(1488..1702)
/label=H1 promoter
/note="human H1 RNA promoter"
primer_bind complement(1548..1567)
/label=H1
/note="Human H1 promoter, forward primer"
promoter 1713..2212
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(1748..1771)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 2145..2164
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 2237..2836
/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(2237..2256)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2733..2753
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
protein_bind 3422..3438
/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)."
promoter complement(3446..3476)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3491..3512
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3629..3646)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3800..4388)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3880..3899)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4559..5419)
/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 5182..5201
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5420..5524)
/gene="bla"
/label=AmpR promoter
primer_bind 5592..5610
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5648..5670)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5770..5789
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5983..6005
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6133..6152
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
1381 agccctcact ccttctctag gcgccggaat tagatcgatc tctcgagaaa aaggaggtga
1441 aagacctgct atctcttgaa tagcaggtct ttcacctccg gggatctgtg gtctcataca
1501 gaacttataa gattcccaaa tccaaagaca tttcacgttt atggtgattt cccagaacac
1561 atagcgacat gcaaatattg cagggcgcca ctcccctgtc cctcacagcc atcttcctgc
1621 cagggcgcac gcgcgctggg tgttcccgcc tagtgacact gggcccgcga ttccttggag
1681 cgggttgatg acgtcagcgt tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca
1741 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
1801 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
1861 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
1921 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
1981 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
2041 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
2101 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
2161 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
2221 cccaagctag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
2281 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
2341 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
2401 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
2461 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
2521 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
2581 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
2641 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
2701 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
2761 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
2821 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
2881 accccatgca tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
2941 agaccccacc tgtaggtttg gcaagctaga gaaccatcag atgtttccag ggtgccccaa
3001 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
3061 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
3121 cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
3181 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
3241 cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg
3301 gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact
3361 ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa
3421 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
3481 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
3541 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
3601 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
3661 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
3721 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
3781 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
3841 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
3901 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
3961 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
4021 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
4081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
4141 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
4201 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
4261 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
4321 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
4381 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
4441 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
4501 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
4561 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
4621 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
4681 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
4741 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
4801 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
4861 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
4921 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
4981 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
5041 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
5101 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
5161 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
5221 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
5281 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
5341 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
5401 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
5461 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
5521 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
5581 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
5641 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
5701 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
5761 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
5821 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
5881 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
5941 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
6001 aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
6061 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
6121 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
6181 gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta aagacaggat
6241 atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta
6301 cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg g
//
