| 质粒名称: | pGL3-U6-sgRNA-EGFP |
|---|---|
| 出品公司： | Addgene |
| 目录编号: | 107721 |
| 存储实验室: | Xingxu Huang |
|
载体骨架基本信息 |
|
| 载体名称: | pGL3-U6-sgRNA-PGK-puromycin |
| 完整质粒大小: | 49827bp |
| 载体类型: | CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
文献引用方法 |
|
| 材料和方法部分: | pGL3-U6-sgRNA-EGFP was a gift from Xingxu Huang (Addgene plasmid # 107721 ; http://n2t.net/addgene:107721 ; RRID:Addgene_107721) |
| 参考文献部分: | Efficient generation of mouse models of human diseases via ABE- and BE-mediated base editing. Liu Z, Lu Z, Yang G, Huang S, Li G, Feng S, Liu Y, Li J, Yu W, Zhang Y, Chen J, Sun Q, Huang X. Nat Commun. 2018 Jun 14;9(1):2338. doi: 10.1038/s41467-018-04768-7. 10.1038/s41467-018-04768-7 PubMed 29904106 |
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 pGL3-U6-sgRNA-EGFP 4927 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION For expression of sgRNA from the U6 promoter with EGFP expression
simultaneously.
ACCESSION .
VERSION .
KEYWORDS pGL3-U6-sgRNA-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4927)
AUTHORS Liu Z, Lu Z, Yang G, Huang S, Li G, Feng S, Liu Y, Li J, Yu W, Zhang
Y, Chen J, Sun Q, Huang X
TITLE Efficient generation of mouse models of human diseases via ABE- and
BE-mediated base editing.
JOURNAL Nat Commun. 2018 Jun 14;9(1):2338. doi: 10.1038/s41467-018-04768-7.
PUBMED 29904106
REFERENCE 2 (bases 1 to 4927)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4927
/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"
misc_RNA 272..347
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
misc_feature 399..516
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 565..1075
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 994..1012
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1084..1803
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1084..1800
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1129..1150)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1390..1409)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1737..1758
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1926..2047
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1963..1982)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2017..2036
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2188..2205)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2359..2947)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2439..2458)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3118..3978)
/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 3741..3760
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3979..4083)
/gene="bla"
/label=AmpR promoter
rep_origin 4110..4565
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4197..4216)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4407..4428
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 4696..4744
/note="synthetic polyadenylation signal"
misc_feature 4758..4849
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 4798..4817
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg tgagaccgag agagggtctc agttttagag ctagaaatag caagttaaaa
301 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttaaagaa
361 ttctcgacct cgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
421 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
481 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
541 gatccacttt ggccgcggct cgagggggtt ggggttgcgc cttttccaag gcagccctgg
601 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
661 tgggactcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
721 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
781 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
841 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
901 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
961 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
1021 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggggga
1081 tccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
1141 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
1201 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
1261 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
1321 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
1381 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
1441 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
1501 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
1561 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
1621 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
1681 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
1741 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
1801 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
1861 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
1921 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
1981 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
2041 tatcttagtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
2101 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
2161 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
2221 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
2281 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
2341 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
2401 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
2461 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
2521 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
2581 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
2641 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
2701 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
2761 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
2821 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
2881 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
2941 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
3001 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
3061 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
3121 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
3181 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
3241 tgctgcaatg ataccgcggg acccacgctc accggctcca gatttatcag caataaacca
3301 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
3361 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
3421 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
3481 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
3541 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
3601 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
3661 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
3721 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
3781 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
3841 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
3901 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
3961 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
4021 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
4081 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
4141 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
4201 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
4261 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
4321 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
4381 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
4441 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
4501 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
4561 aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
4621 ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt
4681 acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
4741 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
4801 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgataggta
4861 ccgattagtg aacggatctc gacggtatcg atcacgagac tagcctcgag cggccgcccc
4921 cttcacc
//
