| 质粒名称: | pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1 |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0002235 |
| 产品用途: | The hCtRNA simultaneously drives the expression of both the targeting DNMT1 gene NickingRNA and pegRNA, harboring dual selection markers of puroR and RFP. |
| 存储实验室: | Zhuobin Liang Lab 梁卓斌课题组 |
|
载体骨架基本信息 |
|
| 载体名称: | pGL3 |
| 表达系统: | 哺乳动物表达 |
| 启动子: | hCtRNA |
| 拷贝数: | 高拷贝 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5alpha |
|
插入基因信息 |
|
| 插入基因名称: | DNMT1-24del-nicking-sgRNA-pegRNA |
| 物种来源: | H.sapiens |
| 密码子优化: | 未优化 |
|
文献引用方法 |
|
| 参考文献部分: |
TRU-PE: A Universal, Trackable Prime Editor Toolkit for Robust Single-and Multi-Locus Genome Engineering
Zhichao Qiu, Keke Sun, Qingwei Zeng, Ziwei Luo, Xinran Liu, Yaping Li, Xiang Lei, Ruilin Zhao, Zhen Zhang, Damia Romero-Moya, Yuan Yang, Xuan Wang, Ehsan Hashemi, Joe Z. Zhang, Xiaolong Wang, Alessandra Giorgetti, Zhuobin Liang
bioRxiv 2026.02.20.706938
|
pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1
价格：元
规格： 2ug质粒（仅用于转化提取） 转染级去内毒素液体即用质粒100μg 转染级去内毒素液体即用质粒500μg 转染级去内毒素液体即用质粒1mg 更大规格-质粒构建-质粒定制-请联系我们
联系方式：I47-825O-882O
买家导航
pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1载体质粒基本信息
pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1质粒图谱载体图谱和pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1载体序列质粒序列多克隆位点信息
pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1质粒载体简介
pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1质粒序列载体序列
LOCUS pGL3-DNMT1-15del-Nicking/PegRNA-PGK-puroR-mRFP1 5695 bp DNA circular SYN 08-NOV-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5695)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 22, 2026 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5695
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 37..61
/label=OL1080
misc_feature 41..115
/label=hCtRNA
primer_bind 56..79
/label=OL464
primer_bind complement(98..115)
/label=OL465
primer_bind complement(101..127)
/label=OL4225
primer_bind complement(101..116)
/label=OL4227
misc_feature 116..136
/label=DNMT1 nickingRNA-del15
primer_bind 121..151
/label=OL4226
misc_RNA 137..212
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind 137..177
/label=OL462
primer_bind complement(193..214)
/label=OL4229
primer_bind complement(195..231)
/label=OL463
primer_bind complement(195..214)
/label=OL4317
misc_feature 213..287
/label=hCtRNA
primer_bind 213..233
/label=OL1080
primer_bind 228..251
/label=OL464
primer_bind complement(270..287)
/label=OL465
primer_bind complement(273..299)
/label=OL4227
primer_bind complement(273..288)
/label=OL4225
misc_feature 288..307
/label=DNMT1 15del-spacer
primer_bind 296..322
/label=OL4228
misc_RNA 308..383
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind 308..348
/label=OL462
primer_bind complement(364..395)
/label=OL4229
primer_bind complement(366..437)
/label=OL4317
primer_bind complement(366..385)
/label=OL463
misc_feature 384..414
/label=RTT
primer_bind 390..433
/label=OL4230
primer_bind complement(394..437)
/label=OL4231
misc_feature 415..428
/label=PBS
primer_bind complement(455..471)
/label=OL1168
misc_feature 472..589
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 638..1148
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1067..1085
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1170..1766
/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(1170..1189)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1666..1686
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 1767..1823
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
RBS 1806..1813
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1824..2501
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(2479..2526)
/label=OL903
primer_bind 2500..2518
/label=OL965
polyA_signal 2648..2769
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2659..2678)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2713..2732
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3017..3034)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3188..3776)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3268..3287)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3947..4807)
/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 4570..4589
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4808..4912)
/gene="bla"
/label=AmpR promoter
rep_origin 4939..5394
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5026..5045)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5236..5257
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 5525..5573
/note="synthetic polyadenylation signal"
misc_feature 5587..5678
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 5587..5603
/label=OL1167
primer_bind 5627..5646
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 agtgaacgga tctcgacggt atcgatcacg agactagcct agagggggta tagctcagtg
61 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc cccctgccct
121 tcagctaaaa taaagggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
181 atcaacttga aaaagtggca ccgagtcggt gcagaggggg tatagctcag tggtagagca
241 tttgactgca gatcaagagg tccccggttc aaatccgggt gccccctgat tcctggtgcc
301 agaaacagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
361 aaaaagtggc accgagtcgg tgcaggagga agctgctaag gactagttct gcccttctgg
421 caccaggatt tttttctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
481 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
541 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
601 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
661 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
721 gcggcgccga ccctgggact cgcacattct tcacgtccgt tcgcagcgtc acccggatct
781 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
841 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
901 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
961 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
1021 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
1081 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
1141 ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg
1201 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
1261 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
1321 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
1381 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
1441 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
1501 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
1561 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
1621 ccgagcgcgc cggggtgccc gccttcctgg aaacctccgc gccccgcaac ctccccttct
1681 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
1741 ggtgcatgac ccgcaagccc ggtgccgcta ctaacttcag cctgctgaag caggctggag
1801 acgtggagga gaaccctgga cctatggcga gtagcgaaga cgttatcaaa gagttcatgc
1861 gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc gaaggtgaag
1921 gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact gaaagttacc aaaggtggtc
1981 cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc aaagcttacg
2041 ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa ggtttcaaat
2101 gggaacgtgt tatgaacttc gaagacggtg gtgttgttac cgttacccag gactcctccc
2161 tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg taccaacttc ccgtccgacg
2221 gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt atgtacccgg
2281 aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac ggtggtcact
2341 acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg ccgggtgctt
2401 acaaaaccga catcaaactg gacatcacct cccacaacga agactacacc atcgttgaac
2461 agtacgaacg tgctgaaggt cgtcactcca ccggtgctta acgcccgccc cacgacccgc
2521 agcgcccgac cgaaaggagc gcacgacccc atgcatcggt acctttaaga ccaatgactt
2581 acaaggcagc tgtagatctt agccactttc tagagtcggg gcggccggcc gcttcgagca
2641 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
2701 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
2761 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
2821 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg
2881 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
2941 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
3001 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
3061 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
3121 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
3181 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
3241 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
3301 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
3361 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
3421 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
3481 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
3541 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
3601 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
3661 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
3721 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
3781 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
3841 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
3901 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
3961 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
4021 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
4081 taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
4141 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
4201 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
4261 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
4321 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
4381 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
4441 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
4501 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
4561 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
4621 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
4681 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
4741 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
4801 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
4861 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
4921 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
4981 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
5041 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
5101 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
5161 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
5221 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
5281 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
5341 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt
5401 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
5461 gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg
5521 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
5581 agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc
5641 tgtccccagt gcaagtgcag gtgccagaac atttctctat cgataggtac cgatt
//
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