| 质粒名称: | psiCHECK2-let-7 8x |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 20931 |
| 存储实验室: | Yukihide Tomari |
|
载体骨架基本信息 |
|
| 载体名称: | psiCHECK-2 |
| 载体出品公司: | promega |
| 空载体大小: | 6273 bp |
| 完整质粒大小: | 6465 bp |
| 载体类型: | 哺乳细胞表达, Insect Expression, Luciferase |
| 载体抗性: | 氨苄霉素 |
| 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: | 8x let-7 target sites |
| 插入大小: | 216 |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | TAAGTACATCAAGAGCTTCG |
|
文献引用方法 |
|
| 材料和方法部分: | psiCHECK2-let-7 8x was a gift from Yukihide Tomari (Addgene plasmid # 20931 ; http://n2t.net/addgene:20931 ; RRID:Addgene_20931) |
| 参考文献部分: | Drosophila Argonaute1 and Argonaute2 Employ Distinct Mechanisms for Translational Repression. Iwasaki S, Kawamata T, Tomari Y. Mol Cell. 2009 Mar 4. ():. 10.1016/j.molcel.2009.02.010 PubMed 19268617 |
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 psiCHECK2-let-7 8x 6465 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS psiCHECK2-let-7 8x
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6465)
AUTHORS Iwasaki S, Kawamata T, Tomari Y
TITLE Drosophila Argonaute1 and Argonaute2 Employ Distinct Mechanisms for
Translational Repression.
JOURNAL Mol Cell. 2009 Mar 4. ():.
PUBMED 19268617
REFERENCE 2 (bases 1 to 6465)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6465
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 237..285
/note="synthetic polyadenylation signal"
promoter 294..1045
/label=HSV TK promoter
/note="herpes simplex virus thymidine kinase promoter"
regulatory 1075..1084
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1081..2733
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/db_xref="GI:42718124"
/protein_id="AAS38485.1"
/translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
RBS 2040..2048
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
polyA_signal 2777..2898
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2788..2807)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2842..2861
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 3031..3135
/gene="bla"
/label=AmpR promoter
CDS 3136..3996
/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(3354..3373)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4167..4755
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4656..4675
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 4884..5241
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4907..4927)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 5092..5227
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5154..5173
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
intron 5311..5443
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 5426..5445
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 5488..5506
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 5510..5519
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 5516..6451
/codon_start=1
/product="Renilla luciferase"
/label=hRluc
/note="human codon-optimized"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
1 tcgagactat acaaggatct acctcagtcg agactataca aggatctacc tcagtcgaga
61 ctatacaagg atctacctca gtcgagacta tacaaggatc tacctcagtc gagactatac
121 aaggatctac ctcagtcgag actatacaag gatctacctc agtcgagact atacaaggat
181 ctacctcagt cgagactata caaggatcta cctcagtcga cgcggccgct ggccgcaata
241 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt
301 cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg cggggggagg
361 gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg gtggccacgg
421 gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg gaggcggggg
481 tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc gccccagagt
541 gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc
601 gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc atccgcacga
661 ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga
721 gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg tcacccggag
781 gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt
841 gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga
901 tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg
961 ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg
1021 agcgaccctg cagcgacccg cttaaaagct tggcattccg gtactgttgg taaagccacc
1081 atggccgatg ctaagaacat taagaagggc cctgctccct tctaccctct ggaggatggc
1141 accgctggcg agcagctgca caaggccatg aagaggtatg ccctggtgcc tggcaccatt
1201 gccttcaccg atgcccacat tgaggtggac atcacctatg ccgagtactt cgagatgtct
1261 gtgcgcctgg ccgaggccat gaagaggtac ggcctgaaca ccaaccaccg catcgtggtg
1321 tgctctgaga actctctgca gttcttcatg ccagtgctgg gcgccctgtt catcggagtg
1381 gccgtggccc ctgctaacga catttacaac gagcgcgagc tgctgaacag catgggcatt
1441 tctcagccta ccgtggtgtt cgtgtctaag aagggcctgc agaagatcct gaacgtgcag
1501 aagaagctgc ctatcatcca gaagatcatc atcatggact ctaagaccga ctaccagggc
1561 ttccagagca tgtacacatt cgtgacatct catctgcctc ctggcttcaa cgagtacgac
1621 ttcgtgccag agtctttcga cagggacaaa accattgccc tgatcatgaa cagctctggg
1681 tctaccggcc tgcctaaggg cgtggccctg cctcatcgca ccgcctgtgt gcgcttctct
1741 cacgcccgcg accctatttt cggcaaccag atcatccccg acaccgctat tctgagcgtg
1801 gtgccattcc accacggctt cggcatgttc accaccctgg gctacctgat ttgcggcttt
1861 cgggtggtgc tgatgtaccg cttcgaggag gagctgttcc tgcgcagcct gcaagactac
1921 aaaattcagt ctgccctgct ggtgccaacc ctgttcagct tcttcgctaa gagcaccctg
1981 atcgacaagt acgacctgtc taacctgcac gagattgcct ctggcggcgc cccactgtct
2041 aaggaggtgg gcgaagccgt ggccaagcgc tttcatctgc caggcatccg ccagggctac
2101 ggcctgaccg agacaaccag cgccattctg attaccccag agggcgacga caagcctggc
2161 gccgtgggca aggtggtgcc attcttcgag gccaaggtgg tggacctgga caccggcaag
2221 accctgggag tgaaccagcg cggcgagctg tgtgtgcgcg gccctatgat tatgtccggc
2281 tacgtgaata accctgaggc cacaaacgcc ctgatcgaca aggacggctg gctgcactct
2341 ggcgacattg cctactggga cgaggacgag cacttcttca tcgtggaccg cctgaagtct
2401 ctgatcaagt acaagggcta ccaggtggcc ccagccgagc tggagtctat cctgctgcag
2461 caccctaaca ttttcgacgc cggagtggcc ggcctgcccg acgacgatgc cggcgagctg
2521 cctgccgccg tcgtcgtgct ggaacacggc aagaccatga ccgagaagga gatcgtggac
2581 tatgtggcca gccaggtgac aaccgccaag aagctgcgcg gcggagtggt gttcgtggac
2641 gaggtgccca agggcctgac cggcaagctg gacgcccgca agatccgcga gatcctgatc
2701 aaggctaaga aaggcggcaa gatcgccgtg taataattct agagtcgggg cggccggccg
2761 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
2821 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
2881 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
2941 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
3001 gatccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
3061 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
3121 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
3181 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
3241 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
3301 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
3361 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
3421 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
3481 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
3541 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
3601 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
3661 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
3721 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
3781 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
3841 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
3901 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
3961 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
4021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
4081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
4141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
4201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
4261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
4321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
4381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
4441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
4501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
4561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
4621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
4681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
4741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
4801 ccttttgctc acatggctcg acagatctgc gcagcaccat ggcctgaaat aacctctgaa
4861 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
4921 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
4981 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
5041 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
5101 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
5161 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
5221 ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagc
5281 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
5341 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
5401 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc
5461 aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag ccaccatggc
5521 ttccaaggtg tacgaccccg agcaacgcaa acgcatgatc actgggcctc agtggtgggc
5581 tcgctgcaag caaatgaacg tgctggactc cttcatcaac tactatgatt ccgagaagca
5641 cgccgagaac gccgtgattt ttctgcatgg taacgctgcc tccagctacc tgtggaggca
5701 cgtcgtgcct cacatcgagc ccgtggctag atgcatcatc cctgatctga tcggaatggg
5761 taagtccggc aagagcggga atggctcata tcgcctcctg gatcactaca agtacctcac
5821 cgcttggttc gagctgctga accttccaaa gaaaatcatc tttgtgggcc acgactgggg
5881 ggcttgtctg gcctttcact actcctacga gcaccaagac aagatcaagg ccatcgtcca
5941 tgctgagagt gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca tcgaggagga
6001 tatcgccctg atcaagagcg aagagggcga gaaaatggtg cttgagaata acttcttcgt
6061 cgagaccatg ctcccaagca agatcatgcg gaaactggag cctgaggagt tcgctgccta
6121 cctggagcca ttcaaggaga agggcgaggt tagacggcct accctctcct ggcctcgcga
6181 gatccctctc gttaagggag gcaagcccga cgtcgtccag attgtccgca actacaacgc
6241 ctaccttcgg gccagcgacg atctgcctaa gatgttcatc gagtccgacc ctgggttctt
6301 ttccaacgct attgtcgagg gagctaagaa gttccctaac accgagttcg tgaaggtgaa
6361 gggcctccac ttcagccagg aggacgctcc agatgaaatg ggtaagtaca tcaagagctt
6421 cgtggagcgc gtgctgaaga acgagcagta attctaggcg atcgc
//
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