| 质粒名称: | pSUPER-retro-puro-shNup93-SpeI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 87331 |
| 存储实验室: | Lei Lu |
|
载体骨架基本信息 |
|
| 载体名称: | modified pSUPER.neo+GFP |
| 载体出品公司: | OligoEngine |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6349 bp |
| 载体修饰: | 我们删除了NheI和XmaI之间的序列尼欧pSUPER.neo公司+GFP并将NheI和XmaI两侧的嘌呤霉素抗性基因连接到载体中。这个产品被修改了尼欧pSUPER.neo公司+GFP用于shRNAs克隆。所有退火的shRNA寡核苷酸均在BglII和XhoI位点间克隆。结扎成功后BglII位点消失。 |
| 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 Nup93 |
| 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-shNup93-SpeI was a gift from Lei Lu (Addgene plasmid # 87331 ; http://n2t.net/addgene:87331 ; RRID:Addgene_87331) |
| 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-shNup93-SpeI 4454 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of shRNA targeting at Nup93.
ACCESSION .
VERSION .
KEYWORDS pSUPER-retro-puro-shNup93-SpeI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4454)
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 4454)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4454
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 276..295
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 363..962
/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(363..382)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 859..879
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(1098..1114)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1098..1114)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(1146..1165)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1147..1165)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1172..1189)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1172..1188)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1181..1203)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 1330..1785
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1417..1436)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1627..1648
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1811..1915
/gene="bla"
/label=AmpR promoter
CDS 1916..2776
/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(2134..2153)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2947..3535
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3436..3455
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3689..3706
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3823..3844
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3859..3889
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3897..3913
/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 3902..3924
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3921..3937
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3921..3937
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 3958..3976
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter complement(4069..4283)
/label=H1 promoter
/note="human H1 RNA promoter"
primer_bind complement(4129..4148)
/label=H1
/note="Human H1 promoter, forward primer"
primer_bind complement(4329..4352)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
ORIGIN
1 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
61 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
121 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
181 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
241 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
301 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagcta
361 gcatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
421 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
481 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
541 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
601 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
661 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
721 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
781 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
841 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
901 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
961 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga
1021 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
1081 ggtttggcaa cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
1141 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
1201 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
1261 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
1321 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
1381 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
1441 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
1501 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
1561 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
1621 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
1681 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
1741 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
1801 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
1861 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
1921 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
1981 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
2041 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
2101 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
2161 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
2221 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
2281 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
2341 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
2401 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
2461 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
2521 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
2581 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
2641 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
2701 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
2761 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
2821 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
2881 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
2941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
3001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
3061 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
3121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
3181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
3241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
3301 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
3361 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
3421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
3481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
3541 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
3601 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
3661 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
3721 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
3781 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
3841 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
3901 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat
3961 taaccctcac taaagggaac aaaagctggt accgggcccc ccctcgagaa aaacacaaag
4021 cctattatca actctcttga agttgataat aggctttgtg ggggatctgt ggtctcatac
4081 agaacttata agattcccaa atccaaagac atttcacgtt tatggtgatt tcccagaaca
4141 catagcgaca tgcaaatatt gcagggcgcc actcccctgt ccctcacagc catcttcctg
4201 ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac tgggcccgcg attccttgga
4261 gcgggttgat gacgtcagcg ttcgaattct accgggtagg ggaggcgctt ttcccaaggc
4321 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
4381 ggcctcgcac acattccaca tccaccggcc ggtaggcgcc aaccggctcc gttctttggt
4441 ggccccttcg cgcc
//
