| 质粒名称: | pRSI9-U6-(sh)-UbiC-TagRFP-2A-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 28289 |
| 存储实验室: | Alex Chenchik, Gus Frangou |
|
载体骨架基本信息 |
|
| 载体名称: | pRSI9-U6-(sh)-UbiC-TagRFP-2A-Puro |
| 载体出品公司: | Cellecta |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7500 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒，RNAi |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | DECIPHER shRNA Expression Vector |
| 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: | BbsI (BpiI)(未破坏) |
| 3' 克隆位点: | BbsI (BpiI)(未破坏) |
| 5' 测序引物 | FwdU6 (CAAGGCTGTTAGAGAGATAATTGGAA) |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pRSI9-U6-(sh)-UbiC-TagRFP-2A-Puro was a gift from Alex Chenchik & Gus Frangou (Addgene plasmid # 28289 ; http://n2t.net/addgene:28289 ; RRID:Addgene_28289) |
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
-UbiC-TagRFP-2A-Puro质粒图谱.png)
载体简介
载体序列
LOCUS pRSI9-U6-(sh)-UbiC-TagRFP-2A-Puro 7500 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION 3rd generation lentiviral plasmid.
ACCESSION .
VERSION .
KEYWORDS pRSI9-U6-(sh)-UbiC-TagRFP-2A-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7500)
TITLE DECIPHER libraries
REFERENCE 2 (bases 1 to 7500)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7500
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 6..232
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 233..413
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 460..585
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1078..1311
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
promoter 1819..2059
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1819..1839
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 1990..2009
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 2194..2311
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2360..2759
/label=UbC promoter
/note="human ubiquitin C promoter"
regulatory 2766..2775
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2772..3482
/codon_start=1
/product="monomeric derivative of red fluorescent protein
from Entacmaea quadricolor (Merzlyak et al., 2007)"
/label=TagRFP
/note="mammalian codon-optimized"
/translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
ARYCDLPSKLGHKLN"
CDS 3489..3542
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 3549..4148
/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(3549..3568)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4045..4065
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 4241..4474
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4546..4667
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4583..4602)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4637..4656
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4707..4842
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4769..4788
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4875..4891)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4875..4891)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4888..4910)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4899..4915
/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(4923..4953)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4968..4989
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5106..5123)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5277..5865)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5357..5376)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6036..6896)
/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 6659..6678
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6897..7001)
/gene="bla"
/label=AmpR promoter
primer_bind 7069..7087
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7125..7147)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7247..7266
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7460..7482
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7474..7491
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7475..7491
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
1801 atggccgccc ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga
1861 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
1921 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
1981 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
2041 tatatcttgt ggaaaggacg aaacaccgga gtcttctttt ttgaagacac ttcgtgcgtt
2101 tggggtttcg gactgtagaa ctctgaacct ctcggtggtc gccgtatcat tagaattctc
2161 gacctcgaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
2221 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
2281 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
2341 actttggccg ccgttcgaag gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
2401 ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc
2461 gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat
2521 cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca
2581 gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg
2641 tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg
2701 tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtc
2761 tagacgccac catggtgtct aagggcgaag agctgattaa ggagaacatg cacatgaagc
2821 tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca
2881 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
2941 tcgccttcga catcctggct accagcttca tgtacggcag cagaaccttc atcaaccaca
3001 cccagggcat ccccgacttc tttaagcagt ccttccctga gggcttcaca tgggagagag
3061 tcaccacata cgaggacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
3121 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga
3181 tgcagaagaa aacactcggc tgggaggcca acaccgagat gctgtacccc gctgacggcg
3241 gcctggaagg cagaagcgac atggccctga agctcgtggg cgggggccac ctgatctgca
3301 acttcaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct
3361 actatgtgga ccacagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc
3421 agcacgaggt ggctgtggcc agatactgcg acctccctag caaactgggg cacaaactta
3481 atggatccga gggcagagga agccttctaa catgcggtga cgtggaggag aatcccggcc
3541 cttccgggat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
3601 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
3661 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
3721 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
3781 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
3841 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
3901 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
3961 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
4021 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
4081 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
4141 gtgcctgaat ctaggtcgac attctacttg gtacctttaa gaccaatgac ttacaaggca
4201 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
4261 caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
4321 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
4381 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
4441 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
4501 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
4561 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
4621 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
4681 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
4741 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
4801 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacc
4861 aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
4921 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
4981 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
5041 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
5101 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
5161 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
5221 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
5281 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
5341 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
5401 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
5461 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
5521 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
5581 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
5641 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
5701 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
5761 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
5821 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
5881 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
5941 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
6001 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
6061 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
6121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
6181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
6241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
6301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
6361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
6421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
6481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
6541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
6601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
6661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
6721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
6781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
6841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
6901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
6961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
7021 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
7081 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
7141 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
7201 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
7261 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
7321 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
7381 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
7441 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
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