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pGL3-BAG3-Nicking/PegRNA-PGK-puroR-mRFP1载体质粒基本信息

质粒名称: pGL3-BAG3-Nicking/PegRNA-PGK-puroR-mRFP1
出品公司: Wekwikgene
目录编号: 0002205
产品用途: The hCtRNA simultaneously drives the expression of both the targeting BAG3 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: BAG3-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-BAG3-Nicking/PegRNA-PGK-puroR-mRFP1质粒图谱载体图谱和pGL3-BAG3-Nicking/PegRNA-PGK-puroR-mRFP1载体序列质粒序列多克隆位点信息

pGL3-BAG3载体图谱

pGL3-BAG3-Nicking/PegRNA-PGK-puroR-mRFP1质粒载体简介

pGL3-BAG3-Nicking/PegRNA-PGK-puroR-mRFP1质粒序列载体序列

LOCUS      pGL3-BAG3-Nicking/PegRNA-PGK-puroR-mRFP1               5676 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 5676)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..23
                     /label=OL1915
     primer_bind     21..55
                     /label=OL2640
     primer_bind     complement(21..44)
                     /label=OL459
     primer_bind     complement(21..44)
                     /label=OL459a
     primer_bind     22..61
                     /label=OL2721
     primer_bind     24..40
                     /label=OL1147
     primer_bind     36..60
                     /label=OL535
     primer_bind     37..61
                     /label=OL1451
     primer_bind     38..115
                     /label=OL409
     misc_feature    41..115
                     /label=hCtRNA
     primer_bind     complement(41..115)
                     /label=OL410
     primer_bind     56..79
                     /label=OL464
     primer_bind     complement(98..116)
                     /label=OL465
     primer_bind     complement(101..115)
                     /label=OL2643
     primer_bind     complement(102..115)
                     /label=OL2641
     misc_feature    116..135
                     /label=BAG3-nicking
     primer_bind     116..131
                     /label=OL301
     primer_bind     135..176
                     /label=OL462
     misc_RNA        136..211
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     136..163
                     /label=OL2722
     primer_bind     136..151
                     /label=OL2642
     primer_bind     complement(194..230)
                     /label=OL463
     misc_feature    212..286
                     /label=hCtRNA
     primer_bind     212..286
                     /label=OL409
     primer_bind     complement(212..286)
                     /label=OL410
     primer_bind     212..232
                     /label=OL2721
     primer_bind     212..232
                     /label=OL1451
     primer_bind     212..231
                     /label=OL535
     primer_bind     212..226
                     /label=OL2640
     primer_bind     227..250
                     /label=OL464
     primer_bind     complement(269..286)
                     /label=OL465
     primer_bind     complement(272..286)
                     /label=OL2643
     primer_bind     complement(273..286)
                     /label=OL2641
     primer_bind     284..302
                     /label=OL209
     misc_feature    287..306
                     /label=BAG3 spacer
     primer_bind     306..334
                     /label=OL2722
     primer_bind     306..322
                     /label=OL2642
     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(365..383)
                     /label=OL463
     misc_feature    383..394
                     /label=RTT
     primer_bind     complement(389..418)
                     /label=OL540
     misc_feature    395..409
                     /label=PBS
     primer_bind     complement(396..409)
                     /label=OL209
     primer_bind     complement(410..433)
                     /label=OL2645
     primer_bind     complement(410..433)
                     /label=OL2723
     primer_bind     complement(416..432)
                     /label=OL1148
     primer_bind     complement(422..441)
                     /label=OL1916
     misc_feature    453..570
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        619..1129
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1048..1066
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1151..1747
                     /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(1151..1170)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1647..1667
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1718..1772
                     /label=OL901
     primer_bind     complement(1733..1755)
                     /label=OL384
     primer_bind     1740..1771
                     /label=OL379
     CDS             1748..1804
                     /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     1748..1804
                     /label=OL866
     primer_bind     complement(1748..1804)
                     /label=OL867
     primer_bind     1785..1828
                     /label=OL902
     primer_bind     complement(1785..1812)
                     /label=OL966
     RBS             1787..1794
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1805..2482
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2460..2507)
                     /label=OL903
     primer_bind     2481..2499
                     /label=OL965
     polyA_signal    2629..2750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2640..2659)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2694..2713
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2998..3015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3169..3757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3249..3268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3928..4788)
                     /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     4551..4570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4789..4893)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4920..5375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5007..5026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5217..5238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5506..5554
                     /note="synthetic polyadenylation signal"
     misc_feature    5568..5659
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5608..5627
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
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 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
      481 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
      541 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
      601 ctttggccgc ggctcgaggg ggttggggtt gcgccttttc caaggcagcc ctgggtttgc
      661 gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggac
      721 tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg
      781 ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc
      841 gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg
      901 ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc
      961 agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt
     1021 agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac
     1081 gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg gggatccacc
     1141 ggagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     1201 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     1261 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     1321 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     1381 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     1441 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     1501 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     1561 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     1621 cgccttcctg gaaacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     1681 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     1741 cggtgccgct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
     1801 acctatggcg agtagcgaag acgttatcaa agagttcatg cgtttcaaag ttcgtatgga
     1861 aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga
     1921 aggtacccag accgctaaac tgaaagttac caaaggtggt ccgctgccgt tcgcttggga
     1981 catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc cggctgacat
     2041 cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt
     2101 cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg gtgagttcat
     2161 ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa
     2221 aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg
     2281 tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg aagttaaaac
     2341 cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg acatcaaact
     2401 ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac gtgctgaagg
     2461 tcgtcactcc accggtgctt aacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     2521 cgcacgaccc catgcatcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     2581 tagccacttt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
     2641 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2701 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2761 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2821 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc
     2881 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2941 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     3001 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3061 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3121 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3181 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3241 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3301 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3361 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3421 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3481 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3541 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3601 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3661 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3721 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3781 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3841 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3901 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3961 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4021 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
     4081 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4141 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4201 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4261 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4321 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4381 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4441 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4501 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4561 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4621 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4681 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4741 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4801 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4861 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4981 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5041 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5101 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5161 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5221 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5281 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5341 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
     5401 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
     5461 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
     5521 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
     5581 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
     5641 ggtgccagaa catttctcta tcgataggta ccgatt
//

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