| 质粒名称: | pGL3-CDKL5-Nicking/PegRNA-PGK-puroR-mRFP1 |
|---|---|
| 出品公司: | Wekwikgene |
| 目录编号: | 0002230 |
| 产品用途: | The hCtRNA simultaneously drives the expression of both the targeting CDKL5 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: | CDKL5-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-CDKL5-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-CDKL5-Nicking/PegRNA-PGK-puroR-mRFP1载体质粒基本信息
pGL3-CDKL5-Nicking/PegRNA-PGK-puroR-mRFP1质粒图谱载体图谱和pGL3-CDKL5-Nicking/PegRNA-PGK-puroR-mRFP1载体序列质粒序列多克隆位点信息
pGL3-CDKL5-Nicking/PegRNA-PGK-puroR-mRFP1质粒载体简介
pGL3-CDKL5-Nicking/PegRNA-PGK-puroR-mRFP1质粒序列载体序列
LOCUS pGL3-CDKL5-Nicking/PegRNA-PGK-puroR-mRFP1 5682 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 5682)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 22, 2026 from SnapGene 6.0.2
https://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5682
/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=OL4191
primer_bind complement(96..116)
/label=OL4193
primer_bind complement(98..115)
/label=OL465
primer_bind 116..155
/label=OL4192
misc_feature 116..135
/label=CDKL5-nickingRNA
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 136..155
/label=OL4194
primer_bind complement(192..212)
/label=4195
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=OL4193
primer_bind complement(267..287)
/label=OL4191
primer_bind complement(269..286)
/label=OL465
primer_bind 287..326
/label=OL4194
misc_feature 287..306
/label=CDKL5-Spacer
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 307..326
/label=OL4192
primer_bind complement(363..437)
/label=4195
primer_bind complement(365..383)
/label=OL463
misc_feature 383..402
/label=RTT
misc_feature 403..415
/label=PBS
misc_feature 459..576
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 625..1135
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1054..1072
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1157..1753
/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(1157..1176)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1653..1673
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 1754..1810
/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 1793..1800
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1811..2488
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(2466..2513)
/label=OL903
primer_bind 2487..2505
/label=OL965
polyA_signal 2635..2756
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2646..2665)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2700..2719
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3004..3021)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3175..3763)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3255..3274)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3934..4794)
/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 4557..4576
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4795..4899)
/gene="bla"
/label=AmpR promoter
rep_origin 4926..5381
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5013..5032)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5223..5244
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 5512..5560
/note="synthetic polyadenylation signal"
misc_feature 5574..5665
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 5614..5633
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 agtgaacgga tctcgacggt atcgatcacg agactagcct agagggggta tagctcagtg
61 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc cccctgcaga
121 accgccactc attcagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
181 tcaacttgaa aaagtggcac cgagtcggtg cagagggggt atagctcagt ggtagagcat
241 ttgactgcag atcaagaggt ccccggttca aatccgggtg ccccctgagg gactcctaga
301 ggactggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
361 aaaagtggca ccgagtcggt gcatattgac acaattcccc aatcctctag gagtcttttt
421 ttctcgacct cgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
481 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
541 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
601 gatccacttt ggccgcggct cgagggggtt ggggttgcgc cttttccaag gcagccctgg
661 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
721 tgggactcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
781 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
841 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
901 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
961 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
1021 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
1081 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggggga
1141 tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
1201 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
1261 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
1321 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
1381 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
1441 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
1501 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
1561 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
1621 ggtgcccgcc ttcctggaaa cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
1681 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
1741 caagcccggt gccgctacta acttcagcct gctgaagcag gctggagacg tggaggagaa
1801 ccctggacct atggcgagta gcgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg
1861 tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc
1921 gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc
1981 ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc
2041 tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat
2101 gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga
2161 gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca
2221 gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct
2281 gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt
2341 taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat
2401 caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc
2461 tgaaggtcgt cactccaccg gtgcttaacg cccgccccac gacccgcagc gcccgaccga
2521 aaggagcgca cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt
2581 agatcttagc cactttctag agtcggggcg gccggccgct tcgagcagac atgataagat
2641 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
2701 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
2761 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
2821 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc gatgcccttg
2881 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
2941 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc
3001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
3061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
3121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
3181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
3241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
3301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
3361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
3421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
3481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
3541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
3601 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
3661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
3721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
3781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
3841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
3901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
3961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
4021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc
4081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
4141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
4201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
4261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
4321 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
4381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
4441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
4501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
4561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
4621 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
4681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
4741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
4801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
4861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
4921 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
4981 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
5041 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
5101 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
5161 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
5221 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
5281 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
5341 atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc cattcaggct
5401 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agcccaagct
5461 accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg caataaaata
5521 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac
5581 gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca
5641 agtgcaggtg ccagaacatt tctctatcga taggtaccga tt
//
