| 质粒名称: | GFP-HuR |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 121162 |
| 存储实验室: | Christine Mayr |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 完整质粒大小: | 6825 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | HuR / ELAVL1 |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-HuR was a gift from Christine Mayr (Addgene plasmid # 121162 ; http://n2t.net/addgene:121162 ; RRID:Addgene_121162) |
| 参考文献部分: | A Membraneless Organelle Associated with the Endoplasmic Reticulum Enables 3'UTR-Mediated Protein-Protein Interactions. Ma W, Mayr C. Cell. 2018 Oct 30. pii: S0092-8674(18)31315-1. doi: 10.1016/j.cell.2018.10.007. 10.1016/j.cell.2018.10.007 PubMed 30449617 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS GFP-HuR 6825 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of GFP tagged HuR.
ACCESSION .
VERSION .
KEYWORDS GFP-HuR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6825)
AUTHORS Ma W, Mayr C
TITLE A Membraneless Organelle Associated with the Endoplasmic Reticulum
Enables 3'UTR-Mediated Protein-Protein Interactions.
JOURNAL Cell. 2018 Oct 30. pii: S0092-8674(18)31315-1. doi:
10.1016/j.cell.2018.10.007.
PUBMED 30449617
REFERENCE 2 (bases 1 to 6825)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6825
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 905..1621
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(950..971)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1211..1230)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1558..1579
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2665..2682)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2671..2895
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2941..3369
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3028..3047)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3238..3259
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3378..3398)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3383..3712
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3563..3698
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3625..3644
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3725..4324
/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(3725..3744)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4221..4241
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4498..4619
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4535..4554)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4589..4608
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4668..4684)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4668..4684)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4681..4703)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4692..4708
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4716..4746)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4761..4782
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4899..4916)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5070..5658)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5150..5169)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5829..6689)
/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 6452..6471
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6690..6794)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
961 ggacggcgac gtaaacggcc acaagttcag cgtgtctggc gagggcgagg gcgatgccac
1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
1081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
1201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
1381 gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
1501 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
1621 gtctaatggt tatgaagacc acatggccga agactgcagg ggtgacatcg ggagaacgaa
1681 tttgatcgtc aactacctcc ctcagaacat gacccaggat gagttacgaa gcctgttcag
1741 cagcattggt gaagttgaat ctgcaaaact tattcgggat aaagtagcag gacacagctt
1801 gggctatggc tttgtgaact acgtgaccgc gaaggatgca gagagagcga tcaacacgct
1861 gaacggcttg aggctccagt caaaaaccat taaggtgtcg tatgctcgcc cgagctcaga
1921 ggtgatcaaa gacgccaact tgtacatcag cgggctcccg cggaccatga cccagaagga
1981 cgtagaagac atgttctctc ggtttgggcg gatcatcaac tcgcgggtcc tcgtggatca
2041 gactacaggt ttgtccagag gggttgcgtt tatccggttt gacaaacggt cggaggcaga
2101 agaggcaatt accagtttca atggtcataa acccccaggt tcctctgagc ccatcacagt
2161 gaagtttgca gccaacccca accagaacaa aaacgtggca ctcctctcgc agctgtacca
2221 ctcgccagcg cgacggttcg gaggccccgt tcaccaccag gcgcagagat tcaggttctc
2281 ccccatgggc gtcgatcaca tgagcgggct ctctggcgtc aacgtgccag gaaacgcctc
2341 ctccggctgg tgcattttca tctacaacct ggggcaggat gccgacgagg ggatcctctg
2401 gcagatgttt gggccgtttg gtgccgtcac caatgtgaaa gtgatccgcg acttcaacac
2461 caacaagtgc aaagggtttg gctttgtgac catgacaaac tatgaagaag ccgcgatggc
2521 catagccagc ctgaacggct accgcctggg ggacaaaatc ttacaggttt ccttcaaaac
2581 caacaagtcc cacaaataac tgcagatatc cagcacagtg gcggccgctc gagtctagag
2641 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
2701 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
2761 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
2821 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
2881 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
2941 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
3001 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
3061 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
3121 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
3181 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
3241 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
3301 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
3361 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
3421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
3541 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
3601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
3661 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
3721 aaccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
3781 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
3841 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
3901 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
3961 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
4021 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
4081 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
4141 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
4201 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
4261 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
4321 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
4381 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
4441 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
4501 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
4561 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
4621 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
4681 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
4741 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
4801 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
4861 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
4921 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
4981 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
5041 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
5101 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
5161 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
5221 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
5281 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
5341 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
5401 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
5461 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
5521 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
5581 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
5641 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
5701 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
5761 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
5821 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
5881 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
5941 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
6001 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
6061 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
6121 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
6181 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
6241 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
6301 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
6361 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
6421 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
6481 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
6541 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
6601 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
6661 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
6721 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
6781 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
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