pGL3-B/N/F-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

相关技术服务：质粒构建    基因合成    质粒大提

联系方式：I47-825O-882O

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pGL3-B/N/F-Nicking/PegRNA-PGK-puroR-mRFP1载体质粒基本信息

质粒名称: pGL3-B/N/F-Nicking/PegRNA-PGK-puroR-mRFP1
出品公司: Wekwikgene
目录编号: 0002208
产品用途: hCtRNA simultaneously drives the expression of NickingRNA and pegRNA targeting three different genes: BAG3, NMB, FANCF, 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: BAG3-nicking-sgRNA-pegRNA, NMB-nicking-sgRNA-pegRNA, FANCF-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-B/N/F-Nicking/PegRNA-PGK-puroR-mRFP1质粒图谱载体图谱和pGL3-B/N/F-Nicking/PegRNA-PGK-puroR-mRFP1载体序列质粒序列多克隆位点信息

pGL3-B载体图谱

pGL3-B/N/F-Nicking/PegRNA-PGK-puroR-mRFP1质粒载体简介

pGL3-B/N/F-Nicking/PegRNA-PGK-puroR-mRFP1质粒序列载体序列

LOCUS       pGL3-B/N/F-Nicking/PegRNA-PGK-puroR-mRFP1              6495 bp DNA     circular SYN 07-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6495)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..23
                     /label=OL1915
     primer_bind     4..23
                     /label=OL1917
     primer_bind     24..40
                     /label=OL1147
     primer_bind     24..40
                     /label=OL1149
     primer_bind     36..60
                     /label=OL535
     misc_feature    41..115
                     /label=hCtRNA
     misc_feature    116..135
                     /label=BAG3-nicking
     misc_RNA        136..211
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    212..286
                     /label=hCtRNA
     primer_bind     212..231
                     /label=OL535
     misc_feature    287..306
                     /label=BAG3 spacer
     misc_RNA        307..382
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    383..394
                     /label=RTT
     misc_feature    395..409
                     /label=PBS
     primer_bind     complement(416..434)
                     /label=OL1148
     primer_bind     428..449
                     /label=OL1149
     primer_bind     433..449
                     /label=OL1147
     primer_bind     445..469
                     /label=OL535
     misc_feature    450..524
                     /label=hCtRNA
     misc_feature    525..544
                     /label=NMB-nicking
     misc_RNA        545..620
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    621..695
                     /label=hCtRNA
     primer_bind     621..640
                     /label=OL535
     misc_feature    696..715
                     /label=NMB spacer
     misc_RNA        716..791
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    792..805
                     /label=RTT
     misc_feature    806..816
                     /label=PBS
     primer_bind     complement(823..841)
                     /label=OL1148
     primer_bind     835..856
                     /label=OL1149
     primer_bind     840..856
                     /label=OL1147
     primer_bind     852..876
                     /label=OL535
     misc_feature    857..931
                     /label=hCtRNA
     misc_feature    857..859
                     /label=5' leader
     misc_feature    932..951
                     /label=FANCF nicking
     misc_RNA        952..1027
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    1028..1102
                     /label=hCtRNA
     primer_bind     1028..1047
                     /label=OL535
     misc_feature    1103..1122
                     /label=FANCF spacer
     misc_RNA        1123..1198
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    1199..1215
                     /label=RT template
     primer_bind     complement(1208..1237)
                     /label=OL550
     misc_feature    1211
                     /label=G to T
     misc_feature    1216..1228
                     /label=PBS
     primer_bind     complement(1222..1240)
                     /label=OL1150
     primer_bind     complement(1235..1251)
                     /label=OL1148
     primer_bind     complement(1241..1261)
                     /label=OL1918
     primer_bind     complement(1241..1260)
                     /label=OL1916
     primer_bind     complement(1255..1271)
                     /label=OL1168
     misc_feature    1272..1389
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     1302..1326
                     /label=U6-shuyuan
     promoter        1438..1948
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1532..1551)
                     /label=OL1068
     CDS             1970..2566
                     /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     2537..2591
                     /label=OL901
     primer_bind     complement(2552..2574)
                     /label=OL384
     primer_bind     2559..2590
                     /label=OL379
     CDS             2567..2623
                     /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"
     primer_bind     2567..2623
                     /label=OL866
     primer_bind     complement(2567..2623)
                     /label=OL867
     primer_bind     2604..2647
                     /label=OL902
     primer_bind     complement(2604..2631)
                     /label=OL966
     RBS             2606..2613
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2624..3301
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(3279..3326)
                     /label=OL903
     primer_bind     3300..3318
                     /label=OL965
     polyA_signal    3448..3569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3451..3470)
                     /label=pCMV5-R
     primer_bind     complement(3459..3478)
                     /label=EBV-R
     primer_bind     3467..3488
                     /label=pFastBac-R
     primer_bind     3513..3532
                     /label=SV40pA-R
     primer_bind     3593..3610
                     /label=pEGFP-C3
     primer_bind     complement(3728..3747)
                     /label=RV4
     rep_origin      complement(3988..4576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4747..5607)
                     /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     complement(4797..4816)
                     /label=Amp-R
     primer_bind     5460..5479
                     /label=Amp-F
     promoter        complement(5608..5712)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5739..6194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6195..6211
                     /label=puc57-F
     polyA_signal    6325..6373
                     /note="synthetic polyadenylation signal"
     misc_feature    6387..6478
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6387..6403
                     /label=OL1167
     primer_bind     6427..6446
                     /label=RV3
     primer_bind     6428..6446
                     /label=pGL3+
ORIGIN
        1 agtgaacgga tctcgacggt atcgatcacg agactagcct agagggggta tagctcagtg
       61 gtagagcatt tgactgcaga tcaagaggtc cccggttcaa atccgggtgc ccccttctcc
      121 ttacctcagg tccgtgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      181 tcaacttgaa aaagtggcac cgagtcggtg cagagggggt atagctcagt ggtagagcat
      241 ttgactgcag atcaagaggt ccccggttca aatccgggtg ccccctactc agccagataa
      301 acagtggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      361 aaaagtggca ccgagtcggt gcacctgtcc acgctgttta tctggctgat ttttttctcg
      421 acctcgagac aagatcacga gactagccta gagggggtat agctcagtgg tagagcattt
      481 gactgcagat caagaggtcc ccggttcaaa tccgggtgcc cccttccttt ccaggtcact
      541 tcatgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      601 aagtggcacc gagtcggtgc agagggggta tagctcagtg gtagagcatt tgactgcaga
      661 tcaagaggtc cccggttcaa atccgggtgc ccccttggtc cctcagggag gtgtggtttt
      721 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
      781 cgagtcggtg cggacagcta cccacacctc cctgagtttt tttctcgacc tcgagacaag
      841 atcacgagac tagcctagag ggggtatagc tcagtggtag agcatttgac tgcagatcaa
      901 gaggtccccg gttcaaatcc gggtgccccc tggggtccca ggtgctgacg tgttttagag
      961 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     1021 tcggtgcaga gggggtatag ctcagtggta gagcatttga ctgcagatca agaggtcccc
     1081 ggttcaaatc cgggtgcccc ctggaatccc ttctgcagca ccgttttaga gctagaaata
     1141 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgcgg
     1201 aaaagcgatc aaggtgctgc agaagggatt tttttctcga cctcgagaca aatggcagta
     1261 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     1321 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     1381 caaaattttc gggtttatta cagggacagc agagatccac tttggccgcg gctcgagggg
     1441 gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg
     1501 gcgtggttcc gggaaacgca gcggcgccga ccctgggact cgcacattct tcacgtccgt
     1561 tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc
     1621 tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga
     1681 agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg
     1741 ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg
     1801 ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc
     1861 cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt
     1921 tgaccgaatc accgacctct ctccccaggg ggatccaccg gagcttacca tgaccgagta
     1981 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     2041 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     2101 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     2161 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     2221 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     2281 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     2341 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     2401 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg aaacctccgc
     2461 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     2521 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccgcta ctaacttcag
     2581 cctgctgaag caggctggag acgtggagga gaaccctgga cctatggcga gtagcgaaga
     2641 cgttatcaaa gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta acggtcacga
     2701 gttcgaaatc gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact
     2761 gaaagttacc aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc cgcagttcca
     2821 gtacggttcc aaagcttacg ttaaacaccc ggctgacatc ccggactacc tgaaactgtc
     2881 cttcccggaa ggtttcaaat gggaacgtgt tatgaacttc gaagacggtg gtgttgttac
     2941 cgttacccag gactcctccc tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg
     3001 taccaacttc ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc
     3061 caccgaacgt atgtacccgg aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa
     3121 actgaaagac ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc
     3181 ggttcagctg ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga
     3241 agactacacc atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta
     3301 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcggt
     3361 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttc tagagtcggg
     3421 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3481 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3541 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3601 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3661 aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     3721 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     3781 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3841 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3901 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3961 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4021 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4081 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4141 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4201 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4261 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4321 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4381 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4441 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4501 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4561 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4621 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4681 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4741 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4801 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4861 tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag atttatcagc
     4921 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4981 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5041 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5101 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5161 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5221 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5281 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5341 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5401 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5461 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5521 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5581 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5641 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5701 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     5761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5881 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5941 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6061 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6181 aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6241 gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta atattaaggt
     6301 acgggaggta cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     6361 ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa
     6421 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat
     6481 cgataggtac cgatt
//

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