| 质粒名称: | pGL3-GBA1-Nicking/PegRNA-PGK-puroR-mRFP1 |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0002225 |
| 产品用途: | The hCtRNA simultaneously drives the expression of both the targeting GBA1 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: | GBA1-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-GBA1-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-GBA1-Nicking/PegRNA-PGK-puroR-mRFP1载体质粒基本信息
pGL3-GBA1-Nicking/PegRNA-PGK-puroR-mRFP1质粒图谱载体图谱和pGL3-GBA1-Nicking/PegRNA-PGK-puroR-mRFP1载体序列质粒序列多克隆位点信息
pGL3-GBA1-Nicking/PegRNA-PGK-puroR-mRFP1质粒载体简介
pGL3-GBA1-Nicking/PegRNA-PGK-puroR-mRFP1质粒序列载体序列
LOCUS pGL3-GBA1-Nicking/PegRNA-PGK-puroR-mRFP1 5679 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 5679)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 22, 2026 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5679
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 24..61
/label=OL2745
misc_feature 41..115
/label=hCtRNA
primer_bind 56..79
/label=OL464
primer_bind complement(96..135)
/label=OL3426
primer_bind complement(96..116)
/label=OL3428
primer_bind complement(98..115)
/label=OL465
primer_bind 116..159
/label=OL3427
misc_feature 116..135
/label=GBA1-nickingRNA
primer_bind 135..159
/label=OL3429
misc_RNA 136..211
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind 136..176
/label=OL462
primer_bind complement(192..211)
/label=OL3430
primer_bind complement(194..230)
/label=OL463
misc_feature 212..286
/label=hCtRNA
primer_bind 212..232
/label=OL2745
primer_bind 227..250
/label=OL464
primer_bind complement(267..306)
/label=OL3428
primer_bind complement(267..290)
/label=OL3426
primer_bind complement(269..286)
/label=OL465
primer_bind 287..330
/label=OL3429
misc_feature 287..306
/label=GBA1 spacer
primer_bind 306..330
/label=OL3427
misc_RNA 307..382
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind 307..347
/label=OL462
primer_bind complement(363..437)
/label=OL3430
primer_bind complement(365..382)
/label=OL463
misc_feature 383..399
/label=RTT
primer_bind 397..412
/label=OL3428
misc_feature 400..412
/label=PBS
misc_feature 456..573
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 622..1132
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1051..1069
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1154..1750
/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(1154..1173)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1650..1670
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 1751..1807
/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 1790..1797
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1808..2485
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(2463..2510)
/label=OL903
primer_bind 2484..2502
/label=OL965
polyA_signal 2632..2753
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2643..2662)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2697..2716
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3001..3018)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3172..3760)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3252..3271)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3931..4791)
/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 4554..4573
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4792..4896)
/gene="bla"
/label=AmpR promoter
rep_origin 4923..5378
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5010..5029)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5220..5241
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 5509..5557
/note="synthetic polyadenylation signal"
misc_feature 5571..5662
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 5611..5630
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 agtgaacgga tctcgacggt atcgatcacg agactagcct agagggggta tagctcagtg
61 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc cccctaccct
121 tacctacact ctctggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
181 tcaacttgaa aaagtggcac cgagtcggtg cagagggggt atagctcagt ggtagagcat
241 ttgactgcag atcaagaggt ccccggttca aatccgggtg ccccctagcc gaccacatgg
301 tacagggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
361 aaaagtggca ccgagtcggt gcttacccta gagcctcctg taccatgtgg tctttttttc
421 tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
481 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
541 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
601 ccactttggc cgcggctcga gggggttggg gttgcgcctt ttccaaggca gccctgggtt
661 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
721 gactcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
781 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
841 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
901 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
961 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
1021 ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg
1081 cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggatcc
1141 accggagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
1201 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
1261 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
1321 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
1381 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
1441 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
1501 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
1561 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
1621 gcccgccttc ctggaaacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
1681 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
1741 gcccggtgcc gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc
1801 tggacctatg gcgagtagcg aagacgttat caaagagttc atgcgtttca aagttcgtat
1861 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
1921 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
1981 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
2041 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
2101 cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
2161 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
2221 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa
2281 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
2341 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
2401 actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga
2461 aggtcgtcac tccaccggtg cttaacgccc gccccacgac ccgcagcgcc cgaccgaaag
2521 gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg cagctgtaga
2581 tcttagccac tttctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
2641 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
2701 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
2761 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
2821 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga
2881 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
2941 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
3001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
3061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
3121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
3181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
3241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
3301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
3361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
3421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
3481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
3541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
3601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
3661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
3721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
3781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
3841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
3901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
3961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
4021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gggacccacg
4081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
4141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
4201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
4261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
4321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
4381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
4441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
4501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
4561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
4621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
4681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
4741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
4801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
4861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
4921 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
4981 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
5041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
5101 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
5161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
5221 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
5281 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
5341 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
5401 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
5461 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
5521 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
5581 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
5641 gcaggtgcca gaacatttct ctatcgatag gtaccgatt
//
