pAcGFP1-C In-Fusion Ready

价格:2500元

联系方式:I47-825O-882O

买家导航

载体基本信息

出品公司: Clontech
载体名称: pAcGFP1-C In-Fusion Ready
质粒类型: 哺乳动物细胞表达载体;荧光报告载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: CMV IE
载体大小: 4711 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: AcGFP1 (N-端)
载体抗性: 卡那霉素
筛选标记: 新霉素(Neomycin)
克隆菌株: DH5α , HB101
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 哺乳动物载体pAcGFP1-C In-Fusion Ready是线性化的载体,可直接用于片段连接。
产品目录号: 634306
稳定性: 稳表达 或 瞬表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pAcGFP1-C In-Fusion Ready载体图谱



pAcGFP1-C In-Fusion Ready 多克隆位点

pAcGFP1-C In-Fusion Ready 载体特征

载体简介

pAcGFP1-C In-Fusion Ready vector is a linearized mammalian expression vector that encodes a Green Fluorescent Protein (GFP) from Aequorea coerulescens. This fluorescent protein coding sequence in this construct has been human codon-optimized for efficient expression and enhanced brightness. AcGFP1 protein has an excitation maximum at 475 nm and an emission maximum at 505 nm. Sequences flanking AcGFP1 have been converted to a Kozak consensus translation initiation site (1) to further increase the translation efficiency in eukaryotic cells. The linearized vector allows direct cloning of PCR products without any need for restriction digestion when using Clontech's
In-Fusion HD Cloning Plus. This is accomplished by the use of a specific 15 nucleotide long sequence within the sense and antisense primers that overlap with the cut ends created by initial digestion of the vector with Sal I and Hind III
The primers that will be used to amplify In-Fusion Ready PCR products require the following 15 nucleotides on their 5’ends:
Sense primer: 5’ AAGGCCTCTGTCGAC followed by sequence of amplification target 3’
Antisense primer: 5’ AGAATTCGCAAGCTT followed by sequence of amplification target 3’
If the gene of interest is added in-frame immediately after the 15 nucleotides mentioned above, this sequence will automatically be in-frame with the AcGFP1 sequence upstream and therefore be expressed as a fusion protein to the C-terminus of AcGFP1.
SV40 polyadenylation signals downstream of the AcGFP1 gene direct proper processing of the 3’ end of the mRNA transcript. In addition, the vector also contains a SV40 origin of replication in mammalian cells expressing the SV40 T-antigen. A neomycin resistance cassette (Neor), containing an SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, andpolyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene, allow selection of stable transformants in eukaryotic cells using G418. A bacterial promoter upstream of the gene expresses kanamycin resistance in E. coli. The pAcGFP1-C In-Fusion Ready Vector also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

Fusions to the C-terminus of AcGFP1 retain the fluorescent properties of the native protein and allow monitoring of the fusion protein localization in vivo. The target gene can be fused into pAcGFP1-C In-Fusion Ready Vector so that it is in-frame with the AcGFP1 coding sequence with no intervening in-frame stop codons. The recombinant AcGFP1 vector can be transfected into mammalian cells using any standard transfection method. Stable transformants can be selected using G418 (2).

Propagation in E. coli
 Suitable host strains: Stellar Competent Cells.
 Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC
 Copy number: ~ 500
 Plasmid incompatibility group: pMB1/ColE1

载体序列

LOCUS       pAcGFP1-C In-Fusion Ready	4711 bp 	DNA	SYN
DEFINITION  pAcGFP1-C In-Fusion Ready
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..4711
                     /organism="pAcGFP1-C In-Fusion Ready"
                     /mol_type="other DNA"
     terminator      40..268
                     /label="SV40_PA_terminator"
     misc_feature    238..257
                     /label="EBV_rev_primer"
     rep_origin      complement(407..713)
                     /label="f1_origin"
     promoter        792..820
                     /label="AmpR_promoter"
     misc_feature    complement(886..906)
                     /label="pBABE_3_primer"
     misc_feature    complement(892..1107)
                     /label="SV40_enhancer"
     promoter        904..1172
                     /label="SV40_promoter"
     rep_origin      1071..1148
                     /label="SV40_origin"
     misc_feature    1133..1152
                     /label="SV40pro_F_primer"
     CDS             1255..2049
                     /label="ORF frame 1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     gene            1258..2046
                     /label="NeoR/KanR"
                     /gene="NeoR/KanR"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     CDS             complement(1564..2100)
                     /label="ORF frame 2"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     terminator      2224..2493
                     /label="TK_PA_terminator"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     rep_origin      2641..3260
                     /label="pBR322_origin"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     promoter        3369..3921
                     /label="CMV_immearly_promoter"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     misc_feature    3424..3711
                     /label="CAG_enhancer"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     misc_feature    3878..3898
                     /label="CMV_fwd_primer"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     promoter        3879..3948
                     /label="CMV_promoter"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
     CDS             3972..34
                     /label="ORF frame 3"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFE
                     DDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKN
                     GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH
                     MIYFGFVTAAAITHGMDELYKSGLKASVEACEFWRIHRI*"
     gene            3975..4688
                     /label="AcGFP1"
                     /gene="AcGFP1"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFE
                     DDGNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKN
                     GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH
                     MIYFGFVTAAAITHGMDELYKSGLKASVEACEFWRIHRI*"
     CDS             complement(4188..46)
                     /label="ORF frame 1"
                     /translation="MISYLDPVDPPEFASFDRGLKSGLVQLIHAVGDGGGGHEAEVDH
                     VIALLVGVLGQGALGGQVVVIGQQHRAIADGGILLVVVGQLHAAILDVVADLEVHLDA
                     ILGLVGHDVHIVGVVVVLHLIAQDVAILLEIGAGQLDAIHQGIALELHLGARLVVAVI
                     LEEDGALLDVALRHGALEEVVLLHVIGVA*"
ORIGIN
    1 AGCTTGCGAA TTCTGGCGGA TCCACCGGAT CTAGATAACT GATCATAATC AGCCATACCA
   61 CATTTGTAGA GGTTTTACTT GCTTTAAAAA ACCTCCCACA CCTCCCCCTG AACCTGAAAC
  121 ATAAAATGAA TGCAATTGTT GTTGTTAACT TGTTTATTGC AGCTTATAAT GGTTACAAAT
  181 AAAGCAATAG CATCACAAAT TTCACAAATA AAGCATTTTT TTCACTGCAT TCTAGTTGTG
  241 GTTTGTCCAA ACTCATCAAT GTATCTTAAC GCGTAAATTG TAAGCGTTAA TATTTTGTTA
  301 AAATTCGCGT TAAATTTTTG TTAAATCAGC TCATTTTTTA ACCAATAGGC CGAAATCGGC
  361 AAAATCCCTT ATAAATCAAA AGAATAGACC GAGATAGGGT TGAGTGTTGT TCCAGTTTGG
  421 AACAAGAGTC CACTATTAAA GAACGTGGAC TCCAACGTCA AAGGGCGAAA AACCGTCTAT
  481 CAGGGCGATG GCCCACTACG TGAACCATCA CCCTAATCAA GTTTTTTGGG GTCGAGGTGC
  541 CGTAAAGCAC TAAATCGGAA CCCTAAAGGG AGCCCCCGAT TTAGAGCTTG ACGGGGAAAG
  601 CCGGCGAACG TGGCGAGAAA GGAAGGGAAG AAAGCGAAAG GAGCGGGCGC TAGGGCGCTG
  661 GCAAGTGTAG CGGTCACGCT GCGCGTAACC ACCACACCCG CCGCGCTTAA TGCGCCGCTA
  721 CAGGGCGCGT CAGGTGGCAC TTTTCGGGGA AATGTGCGCG GAACCCCTAT TTGTTTATTT
  781 TTCTAAATAC ATTCAAATAT GTATCCGCTC ATGAGACAAT AACCCTGATA AATGCTTCAA
  841 TAATATTGAA AAAGGAAGAG TCCTGAGGCG GAAAGAACCA GCTGTGGAAT GTGTGTCAGT
  901 TAGGGTGTGG AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG TATGCAAAGC ATGCATCTCA
  961 ATTAGTCAGC AACCAGGTGT GGAAAGTCCC CAGGCTCCCC AGCAGGCAGA AGTATGCAAA
 1021 GCATGCATCT CAATTAGTCA GCAACCATAG TCCCGCCCCT AACTCCGCCC ATCCCGCCCC
 1081 TAACTCCGCC CAGTTCCGCC CATTCTCCGC CCCATGGCTG ACTAATTTTT TTTATTTATG
 1141 CAGAGGCCGA GGCCGCCTCG GCCTCTGAGC TATTCCAGAA GTAGTGAGGA GGCTTTTTTG
 1201 GAGGCCTAGG CTTTTGCAAA GATCGATCAA GAGACAGGAT GAGGATCGTT TCGCATGATT
 1261 GAACAAGATG GATTGCACGC AGGTTCTCCG GCCGCTTGGG TGGAGAGGCT ATTCGGCTAT
 1321 GACTGGGCAC AACAGACAAT CGGCTGCTCT GATGCCGCCG TGTTCCGGCT GTCAGCGCAG
 1381 GGGCGCCCGG TTCTTTTTGT CAAGACCGAC CTGTCCGGTG CCCTGAATGA ACTGCAAGAC
 1441 GAGGCAGCGC GGCTATCGTG GCTGGCCACG ACGGGCGTTC CTTGCGCAGC TGTGCTCGAC
 1501 GTTGTCACTG AAGCGGGAAG GGACTGGCTG CTATTGGGCG AAGTGCCGGG GCAGGATCTC
 1561 CTGTCATCTC ACCTTGCTCC TGCCGAGAAA GTATCCATCA TGGCTGATGC AATGCGGCGG
 1621 CTGCATACGC TTGATCCGGC TACCTGCCCA TTCGACCACC AAGCGAAACA TCGCATCGAG
 1681 CGAGCACGTA CTCGGATGGA AGCCGGTCTT GTCGATCAGG ATGATCTGGA CGAAGAGCAT
 1741 CAGGGGCTCG CGCCAGCCGA ACTGTTCGCC AGGCTCAAGG CGAGCATGCC CGACGGCGAG
 1801 GATCTCGTCG TGACCCATGG CGATGCCTGC TTGCCGAATA TCATGGTGGA AAATGGCCGC
 1861 TTTTCTGGAT TCATCGACTG TGGCCGGCTG GGTGTGGCGG ACCGCTATCA GGACATAGCG
 1921 TTGGCTACCC GTGATATTGC TGAAGAGCTT GGCGGCGAAT GGGCTGACCG CTTCCTCGTG
 1981 CTTTACGGTA TCGCCGCTCC CGATTCGCAG CGCATCGCCT TCTATCGCCT TCTTGACGAG
 2041 TTCTTCTGAG CGGGACTCTG GGGTTCGAAA TGACCGACCA AGCGACGCCC AACCTGCCAT
 2101 CACGAGATTT CGATTCCACC GCCGCCTTCT ATGAAAGGTT GGGCTTCGGA ATCGTTTTCC
 2161 GGGACGCCGG CTGGATGATC CTCCAGCGCG GGGATCTCAT GCTGGAGTTC TTCGCCCACC
 2221 CTAGGGGGAG GCTAACTGAA ACACGGAAGG AGACAATACC GGAAGGAACC CGCGCTATGA
 2281 CGGCAATAAA AAGACAGAAT AAAACGCACG GTGTTGGGTC GTTTGTTCAT AAACGCGGGG
 2341 TTCGGTCCCA GGGCTGGCAC TCTGTCGATA CCCCACCGAG ACCCCATTGG GGCCAATACG
 2401 CCCGCGTTTC TTCCTTTTCC CCACCCCACC CCCCAAGTTC GGGTGAAGGC CCAGGGCTCG
 2461 CAGCCAACGT CGGGGCGGCA GGCCCTGCCA TAGCCTCAGG TTACTCATAT ATACTTTAGA
 2521 TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT TTTGATAATC
 2581 TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA
 2641 AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA
 2701 AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC
 2761 CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTTCTTCTA GTGTAGCCGT
 2821 AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC
 2881 TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC
 2941 GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA
 3001 GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG
 3061 CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG
 3121 GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT
 3181 TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT
 3241 GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC
 3301 ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC GCCATGCATT
 3361 AGTTATTAAT AGTAATCAAT TACGGGGTCA TTAGTTCATA GCCCATATAT GGAGTTCCGC
 3421 GTTACATAAC TTACGGTAAA TGGCCCGCCT GGCTGACCGC CCAACGACCC CCGCCCATTG
 3481 ACGTCAATAA TGACGTATGT TCCCATAGTA ACGCCAATAG GGACTTTCCA TTGACGTCAA
 3541 TGGGTGGAGT ATTTACGGTA AACTGCCCAC TTGGCAGTAC ATCAAGTGTA TCATATGCCA
 3601 AGTACGCCCC CTATTGACGT CAATGACGGT AAATGGCCCG CCTGGCATTA TGCCCAGTAC
 3661 ATGACCTTAT GGGACTTTCC TACTTGGCAG TACATCTACG TATTAGTCAT CGCTATTACC
 3721 ATGGTGATGC GGTTTTGGCA GTACATCAAT GGGCGTGGAT AGCGGTTTGA CTCACGGGGA
 3781 TTTCCAAGTC TCCACCCCAT TGACGTCAAT GGGAGTTTGT TTTGGCACCA AAATCAACGG
 3841 GACTTTCCAA AATGTCGTAA CAACTCCGCC CCATTGACGC AAATGGGCGG TAGGCGTGTA
 3901 CGGTGGGAGG TCTATATAAG CAGAGCTGGT TTAGTGAACC GTCAGATCCG CTAGCGCTAC
 3961 CGGTCGCCAC CATGGTGAGC AAGGGCGCCG AGCTGTTCAC CGGCATCGTG CCCATCCTGA
 4021 TCGAGCTGAA TGGCGATGTG AATGGCCACA AGTTCAGCGT GAGCGGCGAG GGCGAGGGCG
 4081 ATGCCACCTA CGGCAAGCTG ACCCTGAAGT TCATCTGCAC CACCGGCAAG CTGCCTGTGC
 4141 CCTGGCCCAC CCTGGTGACC ACCCTGAGCT ACGGCGTGCA GTGCTTCTCA CGCTACCCCG
 4201 ATCACATGAA GCAGCACGAC TTCTTCAAGA GCGCCATGCC TGAGGGCTAC ATCCAGGAGC
 4261 GCACCATCTT CTTCGAGGAT GACGGCAACT ACAAGTCGCG CGCCGAGGTG AAGTTCGAGG
 4321 GCGATACCCT GGTGAATCGC ATCGAGCTGA CCGGCACCGA TTTCAAGGAG GATGGCAACA
 4381 TCCTGGGCAA TAAGATGGAG TACAACTACA ACGCCCACAA TGTGTACATC ATGACCGACA
 4441 AGGCCAAGAA TGGCATCAAG GTGAACTTCA AGATCCGCCA CAACATCGAG GATGGCAGCG
 4501 TGCAGCTGGC CGACCACTAC CAGCAGAATA CCCCCATCGG CGATGGCCCT GTGCTGCTGC
 4561 CCGATAACCA CTACCTGTCC ACCCAGAGCG CCCTGTCCAA GGACCCCAAC GAGAAGCGCG
 4621 ATCACATGAT CTACTTCGGC TTCGTGACCG CCGCCGCCAT CACCCACGGC ATGGATGAGC
 4681 TGTACAAGTC CGGACTTAAG GCCTCTGTCG A
//

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