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pGL3-ERCC6-Nicking/PegRNA-PGK-puroR-mRFP1载体质粒基本信息

质粒名称: pGL3-ERCC6-Nicking/PegRNA-PGK-puroR-mRFP1
出品公司: Wekwikgene
目录编号: 0002202
产品用途: The hCtRNA simultaneously drives the expression of both the targeting ERCC6 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: ERCC6-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-ERCC6-Nicking/PegRNA-PGK-puroR-mRFP1质粒图谱载体图谱和pGL3-ERCC6-Nicking/PegRNA-PGK-puroR-mRFP1载体序列质粒序列多克隆位点信息

pGL3-ERCC6-Nicking载体图谱

pGL3-ERCC6-Nicking/PegRNA-PGK-puroR-mRFP1质粒载体简介

pGL3-ERCC6-Nicking/PegRNA-PGK-puroR-mRFP1质粒序列载体序列

LOCUS       pGL3-ERCC6-Nicking/PegRNA-PGK-puroR-mRFP1                5682 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 5682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 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     21..55
                     /label=OL2640
     primer_bind     22..61
                     /label=OL2721
     primer_bind     24..40
                     /label=OL1149
     misc_feature    41..115
                     /label=hCtRNA
     primer_bind     56..79
                     /label=OL464
     primer_bind     complement(98..115)
                     /label=OL465
     primer_bind     complement(101..135)
                     /label=OL2651
     primer_bind     complement(101..115)
                     /label=OL2653
     primer_bind     116..164
                     /label=OL2726
     primer_bind     116..151
                     /label=OL2652
     misc_feature    116..135
                     /label=ERCC6 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     complement(194..230)
                     /label=OL463
     misc_feature    212..286
                     /label=hCtRNA
     primer_bind     212..232
                     /label=OL2721
     primer_bind     212..226
                     /label=OL2640
     primer_bind     227..250
                     /label=OL464
     primer_bind     complement(269..287)
                     /label=OL465
     primer_bind     complement(272..306)
                     /label=OL2653
     primer_bind     complement(272..286)
                     /label=OL2651
     primer_bind     287..310
                     /label=OL2654
     misc_feature    287..306
                     /label=ERCC6 spacer
     primer_bind     287..301
                     /label=OL2655
     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..335
                     /label=OL2726
     primer_bind     307..322
                     /label=OL2652
     primer_bind     complement(365..383)
                     /label=OL463
     misc_feature    383..397
                     /label=RTT
     primer_bind     complement(393..439)
                     /label=OL2727
     misc_feature    397
                     /label=a to g
     misc_feature    398..415
                     /label=PBS
     primer_bind     complement(398..414)
                     /label=OL2654
     primer_bind     complement(400..439)
                     /label=OL2655
     primer_bind     complement(408..427)
                     /label=OL2756
     primer_bind     421..440
                     /label=OL458
     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"
     primer_bind     1724..1778
                     /label=OL901
     primer_bind     complement(1739..1761)
                     /label=OL384
     primer_bind     1746..1777
                     /label=OL379
     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"
     primer_bind     1754..1810
                     /label=OL866
     primer_bind     complement(1754..1810)
                     /label=OL867
     primer_bind     1791..1834
                     /label=OL902
     primer_bind     complement(1791..1818)
                     /label=OL966
     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 cccctgtcag
      121 tgattagtgg aaatggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      181 tcaacttgaa aaagtggcac cgagtcggtg cagagggggt atagctcagt ggtagagcat
      241 ttgactgcag atcaagaggt ccccggttca aatccgggtg cccccttcaa tgctttcatc
      301 accagagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      361 aaaagtggca ccgagtcggt gcacttctat gccatccggt gatgaaagca ttgatttttt
      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
//

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