| 质粒名称: | pShuttle-CMV-GFP U6promoter |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 128049 |
| 存储实验室: | Oskar Laur |
|
载体骨架基本信息 |
|
| 载体名称: | pShuttle |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,腺病毒 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 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: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pShuttle-CMV-GFP U6promoter was a gift from Oskar Laur (Addgene plasmid # 128049 ; http://n2t.net/addgene:128049 ; RRID:Addgene_128049) |
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 pShuttle-CMV-GFP U6promoter 8233 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Adenoviral shuttle vector for shRNA expression from U6 promoter with
GFP marker.
ACCESSION .
VERSION .
KEYWORDS pShuttle-CMV-GFP U6promoter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8233)
TITLE Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE 2 (bases 1 to 8233)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8233
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
enhancer 363..666
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 667..870
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 821..841
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 909..918
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 915..1631
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 915..1631
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(960..981)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1221..1240)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1568..1589
/label=EGFP-C
/note="EGFP, forward primer"
repeat_region 4827..4929
/label=ITR
/note="inverted terminal repeat of human adenovirus
serotype 5"
primer_bind complement(5018..5035)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5189..5777)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5269..5288)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 6609..7403
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6663..6682)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 7273..7292
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
rep_origin complement(7451..7831)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7513..7534)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7725..7744
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 7869..7971
/label=ITR
/note="inverted terminal repeat of human adenovirus
serotype 5"
misc_signal 8059..8209
/label=Ad5 Psi
/note="packaging signal for adenovirus serotype 5"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg gaagataacc tgcttcaaca cgaatgttga agcaggttat cttccttttt
301 tctagagatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
361 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
421 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
481 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
541 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
601 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
661 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
721 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
781 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
841 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
901 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
961 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
1021 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
1081 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
1141 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
1201 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
1261 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
1321 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
1381 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
1441 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
1501 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
1561 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
1621 agctgtacaa gtccggactc agatctgggc gtggttaagg gtgggaaaga atatataagg
1681 tgggggtctt atgtagtttt gtatctgttt tgcagcagcc gccgccgcca tgagcaccaa
1741 ctcgtttgat ggaagcattg tgagctcata tttgacaacg cgcatgcccc catgggccgg
1801 ggtgcgtcag aatgtgatgg gctccagcat tgatggtcgc cccgtcctgc ccgcaaactc
1861 tactaccttg acctacgaga ccgtgtctgg aacgccgttg gagactgcag cctccgccgc
1921 cgcttcagcc gctgcagcca ccgcccgcgg gattgtgact gactttgctt tcctgagccc
1981 gcttgcaagc agtgcagctt cccgttcatc cgcccgcgat gacaagttga cggctctttt
2041 ggcacaattg gattctttga cccgggaact taatgtcgtt tctcagcagc tgttggatct
2101 gcgccagcag gtttctgccc tgaaggcttc ctcccctccc aatgcggttt aaaacataaa
2161 taaaaaacca gactctgttt ggatttggat caagcaagtg tcttgctgtc tttatttagg
2221 ggttttgcgc gcgcggtagg cccgggacca gcggtctcgg tcgttgaggg tcctgtgtat
2281 tttttccagg acgtggtaaa ggtgactctg gatgttcaga tacatgggca taagcccgtc
2341 tctggggtgg aggtagcacc actgcagagc ttcatgctgc ggggtggtgt tgtagatgat
2401 ccagtcgtag caggagcgct gggcgtggtg cctaaaaatg tctttcagta gcaagctgat
2461 tgccaggggc aggcccttgg tgtaagtgtt tacaaagcgg ttaagctggg atgggtgcat
2521 acgtggggat atgagatgca tcttggactg tatttttagg ttggctatgt tcccagccat
2581 atccctccgg ggattcatgt tgtgcagaac caccagcaca gtgtatccgg tgcacttggg
2641 aaatttgtca tgtagcttag aaggaaatgc gtggaagaac ttggagacgc ccttgtgacc
2701 tccaagattt tccatgcatt cgtccataat gatggcaatg ggcccacggg cggcggcctg
2761 ggcgaagata tttctgggat cactaacgtc atagttgtgt tccaggatga gatcgtcata
2821 ggccattttt acaaagcgcg ggcggagggt gccagactgc ggtataatgg ttccatccgg
2881 cccaggggcg tagttaccct cacagatttg catttcccac gctttgagtt cagatggggg
2941 gatcatgtct acctgcgggg cgatgaagaa aacggtttcc ggggtagggg agatcagctg
3001 ggaagaaagc aggttcctga gcagctgcga cttaccgcag ccggtgggcc cgtaaatcac
3061 acctattacc ggctgcaact ggtagttaag agagctgcag ctgccgtcat ccctgagcag
3121 gggggccact tcgttaagca tgtccctgac tcgcatgttt tccctgacca aatccgccag
3181 aaggcgctcg ccgcccagcg atagcagttc ttgcaaggaa gcaaagtttt tcaacggttt
3241 gagaccgtcc gccgtaggca tgcttttgag cgtttgacca agcagttcca ggcggtccca
3301 cagctcggtc acctgctcta cggcatctcg atccagcata tctcctcgtt tcgcgggttg
3361 gggcggcttt cgctgtacgg cagtagtcgg tgctcgtcca gacgggccag ggtcatgtct
3421 ttccacgggc gcagggtcct cgtcagcgta gtctgggtca cggtgaaggg gtgcgctccg
3481 ggctgcgcgc tggccagggt gcgcttgagg ctggtcctgc tggtgctgaa gcgctgccgg
3541 tcttcgccct gcgcgtcggc caggtagcat ttgaccatgg tgtcatagtc cagcccctcc
3601 gcggcgtggc ccttggcgcg cagcttgccc ttggaggagg cgccgcacga ggggcagtgc
3661 agacttttga gggcgtagag cttgggcgcg agaaataccg attccgggga gtaggcatcc
3721 gcgccgcagg ccccgcagac ggtctcgcat tccacgagcc aggtgagctc tggccgttcg
3781 gggtcaaaaa ccaggtttcc cccatgcttt ttgatgcgtt tcttacctct ggtttccatg
3841 agccggtgtc cacgctcggt gacgaaaagg ctgtccgtgt ccccgtatac agacttgaga
3901 gggagtttaa acgaattcaa tagcttgttg catgggcggc gatataaaat gcaaggtgct
3961 gctcaaaaaa tcaggcaaag cctcgcgcaa aaaagaaagc acatcgtagt catgctcatg
4021 cagataaagg caggtaagct ccggaaccac cacagaaaaa gacaccattt ttctctcaaa
4081 catgtctgcg ggtttctgca taaacacaaa ataaaataac aaaaaaacat ttaaacatta
4141 gaagcctgtc ttacaacagg aaaaacaacc cttataagca taagacggac tacggccatg
4201 ccggcgtgac cgtaaaaaaa ctggtcaccg tgattaaaaa gcaccaccga cagctcctcg
4261 gtcatgtccg gagtcataat gtaagactcg gtaaacacat caggttgatt cacatcggtc
4321 agtgctaaaa agcgaccgaa atagcccggg ggaatacata cccgcaggcg tagagacaac
4381 attacagccc ccataggagg tataacaaaa ttaataggag agaaaaacac ataaacacct
4441 gaaaaaccct cctgcctagg caaaatagca ccctcccgct ccagaacaac atacagcgct
4501 tccacagcgg cagccataac agtcagcctt accagtaaaa aagaaaacct attaaaaaaa
4561 caccactcga cacggcacca gctcaatcag tcacagtgta aaaaagggcc aagtgcagag
4621 cgagtatata taggactaaa aaatgacgta acggttaaag tccacaaaaa acacccagaa
4681 aaccgcacgc gaacctacgc ccagaaacga aagccaaaaa acccacaact tcctcaaatc
4741 gtcacttccg ttttcccacg ttacgtcact tcccatttta agaaaactac aattcccaac
4801 acatacaagt tactccgccc taaaacctac gtcacccgcc ccgttcccac gccccgcgcc
4861 acgtcacaaa ctccaccccc tcattatcat attggcttca atccaaaata aggtatatta
4921 ttgatgatgt taattaacat gcatggatcc atatgcggtg tgaaataccg cacagatgcg
4981 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
5041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
5101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
5161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
5221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
5281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
5761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
5821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
5881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
5941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
6001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
6061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
6121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
6181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
6241 tgcgcaacgt tgttgccatt gctgcagcca tgagattatc aaaaaggatc ttcacctaga
6301 tccttttcac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
6361 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
6421 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
6481 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
6541 tgccgccaag gatctgatgg cgcaggggat caagctctga tcaagagaca ggatgaggat
6601 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
6661 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
6721 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
6781 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
6841 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
6901 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
6961 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
7021 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
7081 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
7141 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
7201 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
7261 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
7321 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
7381 gccttcttga cgagttcttc tgaattttgt taaaattttt gttaaatcag ctcatttttt
7441 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
7501 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
7561 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
7621 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
7681 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
7741 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
7801 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggatc gaattaattc
7861 ttaattaaca tcatcaataa tataccttat tttggattga agccaatatg ataatgaggg
7921 ggtggagttt gtgacgtggc gcggggcgtg ggaacggggc gggtgacgta gtagtgtggc
7981 ggaagtgtga tgttgcaagt gtggcggaac acatgtaagc gacggatgtg gcaaaagtga
8041 cgtttttggt gtgcgccggt gtacacagga agtgacaatt ttcgcgcggt tttaggcgga
8101 tgttgtagta aatttgggcg taaccgagta agatttggcc attttcgcgg gaaaactgaa
8161 taagaggaag tgaaatctga ataattttgt gttactcata gcgcgtaata ctggtaccaa
8221 ggtcgggcag gaa
//
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