| 质粒名称: | NRCoC |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74300 |
| 存储实验室: | Rita Shiang |
|
载体骨架基本信息 |
|
| 载体名称: | pS65T-C1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4728 bp |
| 完整质粒大小: | 6030 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | treacle ribosome biogenesis factor 1,Tcof1 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1302 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | GFP（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | PstI（销毁后未知） |
| 5' 测序引物 | GFP-F |
| 3' 测序引物 | pEGFP-R |
|
文献引用方法 |
|
| 材料和方法部分: | NRCoC was a gift from Rita Shiang (Addgene plasmid # 74300 ; http://n2t.net/addgene:74300 ; RRID:Addgene_74300) |
| 参考文献部分: | The Treacher Collins syndrome (TCOF1) gene product, treacle, is targeted to the nucleolus by signals in its C-terminus. Winokur ST, Shiang R. Hum Mol Genet. 1998 Nov;7(12):1947-52. ddb229 [pii] PubMed 9811939 |
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 NRCoC 8741 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of full-length treacle fused to GFP.
ACCESSION .
VERSION .
KEYWORDS NRCoC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8741)
AUTHORS Winokur ST, Shiang R
TITLE The Treacher Collins syndrome (TCOF1) gene product, treacle, is
targeted to the nucleolus by signals in its C-terminus.
JOURNAL Hum Mol Genet. 1998 Nov;7(12):1947-52.
PUBMED 9811939
REFERENCE 2 (bases 1 to 8741)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8741
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..711
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MGKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELY"
primer_bind complement(31..59)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 654..674
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
primer_bind complement(4735..4751)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4735..4751)
/label=pBluescriptSK
/note="For pBluescript vector"
polyA_signal 4917..5038
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4954..4973)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5008..5027
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(5045..5500)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5182..5203)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5394..5413
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5527..5631
/gene="bla"
/label=AmpR promoter
promoter 5633..5990
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(5656..5676)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 5841..5976
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5903..5922
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 6025..6819
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6079..6098)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6689..6708
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(7007..7026)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 7051..7098
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 7427..8015
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7916..7935
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 8190..8493
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 8494..8697
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 8648..8668
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 8736..4
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
ORIGIN
1 atgggtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
61 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
121 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
181 gtcactactt ttacgtatgg tgttcaatgc ttttcaagat acccagatca tatgaaacgg
241 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
301 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
361 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
421 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
481 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
541 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
601 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
661 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caagtccgga
721 ctcagatctc gaggtgaatc ccctggccag atctctggca tggccgaggc caggaagcgg
781 cgggagctgc ttcccctcat ttaccatcat ctgttgcaag caggctacgt ccgcgcggcg
841 cgggaagtaa aggagcagag cggccagaag agtttcctga ctcagcccgt cacccttctg
901 gacatctata cacactggca acagacctca gagcttggcc agaagcagaa ggcagaggat
961 gatgagaccc ttcaggctaa gaagtctcga gtgtcggatc ctgttagcag ctcagagagc
1021 tcggatcagg agaaggaaga ggaggcagca accgaaaggg ccaaagccac cccaagaccg
1081 acacctgtca attctgcaac cgcagctttg ccatcaaaag taaaagaaaa gggaaagacc
1141 aagacagcca acaagacggt gaactctgtg tcgcaccctg ggtccggaaa gacggtggtc
1201 cacctgctct ctgggaagtc acccaaaaag tcagcagagc ccttggcaaa cactgtcttg
1261 gcctcagaaa ctgaggagga gggcaatgcc caagccctcg gacccactgc caagtctgga
1321 acggtgtcag cgggccaagg cagcagttcc agtgaagatt cctccatctc aagcgatgag
1381 acagatgtcg aggtgaaatc tccagcaaaa ccagcccagg ccaaagcttc agcagcccct
1441 gccaaggatc ctccagcaag aacagcccca ggccctacca agttagggaa tgtggcgccc
1501 acacctgcta aaccagccag ggcggcagcg gcagctgctg ctgctgctgt ggctgctgct
1561 gctgctgcag cagcagaaga gtctgagagc agtgaggagg actcagacag tgaggacgag
1621 gcccctgctg gtctgcccag ccaggtaaaa gcctctggaa aaggtcccca tgtcagagcc
1681 gactcggtgt ctgccaaggg gatctctggg aaagggccca tcttagcaac cccagggaag
1741 actgggcctg cagccaccca ggccaaggca gaaaggccag agaaggactc ggaaaccagc
1801 agtgaggacg attctgatag tgaggatgaa atgccagtca ctgtgaatac tcctcaggca
1861 aggacttctg ggaagagccc tcgggccaga ggtacctcag cccccgccaa ggagtcatcc
1921 cagaaagggg ctcctgcagt cacccctgga aaggcaaggc ctgtggcagc ccaggcaggg
1981 aaaccagaag ccaagagcag tgaggagtca gagagtgaca gtggggagac accagctgct
2041 gcgactctga ccacgagtcc tgccaaggtg aaacctttgg ggaagagctc ccaggtcaga
2101 cctgtttcca ccgtcacccc ggggtcatcg ggaaaaggtg ccaacctgcc ctgccctggg
2161 aaggtggggt cagcagctct cagggtccaa atggtaaaga aagaagatgt ctcggagagc
2221 agcagtgcag agctggacag tgacgggcct gggagcccag ccaaggcaaa ggcctccctt
2281 gcgctccctc agaaggtgag gcctgtggcc acccaggtca agactgacag gggcaaaggc
2341 cactcaggga gcagtgagga gtcatctgac agcgaagagg aggcagcacc agcagcctct
2401 gctgctcagg ctaagccagc tctggaaaag cagatgaaag cttcctctag gaaaggcacg
2461 cctgcatccg caacaggagc gagcacctcg tcccattgta aggcaggagc ggtgacctct
2521 tcagccagcc tgtcatcccc agctctggcc aagggcaccc agaggtcaga tgtggactct
2581 tccagtgagt ctgagtcaga aggagctgct cccagcaccc ccagggtaca ggggaagtct
2641 gggggcaagg gcctccaagg gaaagctgcc ttggggcaag gggtggcccc agtgcacact
2701 cagaagacag ggccttcggt caaagctatg gctcaggaag actcagagag cctcgaggag
2761 gactccagca gtgaggaaga ggatgagacc ccagcacagg ccacgccctt ggggagactt
2821 cctcaggcca aagccaaccc acctcccact aagacacctc cagcgtctgc atctggaaaa
2881 gctgtggctg ctccaaccaa gggaaaacca cctgttccga acagcaccgt ctctgcaagg
2941 ggccagcggt ctgtgccagc cgcgggaaaa gcaggggccc cagcaaccca agcccagaag
3001 ggtcccgtgg ctggcacagg ggaggactca gagagcagca gtaaagagga gtctgacagt
3061 gaagaagaga cgccagccca gataaaacct gtggggaaga cctctcaggt cagagctgcc
3121 tcagcccctg ccaaggagtc tcctaaaaaa ggagcccatc caggaacccc cggcaagacg
3181 ggatcttcag ctacccaggc ccagccaggg aagacagagg actcagacag cagtagtgaa
3241 gagtctgaca gtgacacaga gatgccatca gcccaggatg ctatctccca gcctgccaga
3301 ggcaaagcgt caggtccagc gtccccagag aagagcatag aagggtcctc agagagcagt
3361 gatgaggatc tgccctccgg ccaggcgatt aaatcccctc cagtttctgt caaccgtaac
3421 agtagtccag ctgtcccagc tcctacccca gaaggagtcc aggctgtgaa caccacaaag
3481 aaggcctcag gcaccactgc ccagagctcc tcctctgaga gtgaggacgg ggacgaggac
3541 ttgattcctg ccacacaacc ctccacctat gctctcagaa ccagtgtgac gacgcccgca
3601 gccctctcac gagcagcttc ccaacccagc aagagtgagc agtctagccg gatgccaaaa
3661 ggcaagaaag caaaggcggc ggcgtccgct cagaccagca gtgccgtgga aacactcccc
3721 atgatgcctc cccagagcgc acccatccag cccaaagcca ccaacaagct cgggaaatcc
3781 aagctccctg agaagcagca gcttgcccca ggctacccca aggcccccag gagctcggag
3841 gacagcagtg acacttcttc cgaggacgag gaggatgcca agagacccca gatgcccaag
3901 tcggcccaca ggctggatcc agacccttcc cagaaggaaa ctgtggtaga ggagacccct
3961 acagaatcca gcgaagatga gatggtggcc ccctcacagt ctctcctctc aggttacatg
4021 actcccggct tgactgtggc taattcccag gcttcaaaag ctactcctag gccagactcc
4081 aactccttag cttcttctgc cccggccacc aaagacaacc cggatggcaa gcagaagtca
4141 aaatcccaac acgcagcaga caccgcactc cctaaaaccg gtaggaaaga ggcctcctcg
4201 ggctccacgc ctcagaagcc caagaagctc aagaagagca cctcaagctc gccggccccc
4261 acacagacac tgccgaacag catcacccag cgcctcctgg agcaggcctg gcccctgagc
4321 gaggcgcagg tgcaggcctc tgtggtgaag gtcctgacag agcttctgga gcaggaacgg
4381 ctgaaggcca cagaggccat caaggagagt gggaagaaga gccagaagcg gaagttatca
4441 ggggacctag aagctggggc cccaaagaac aagaagaaga aggagcagcc agtgcccagg
4501 gcaagcgctg tttccccaga aaaggctccc atgacttcca aggcaaaatc aaagcttgac
4561 aaagggagtg ctggtggcaa ggggaagggg tctcctggcc cccaaggagc caaggagaag
4621 cccgacggcg agttgctggg gataaagctt gagagtggcg agcagagcga cccgaagagc
4681 aagtcgaaaa agaagaaatc cctcaagctc gtgccgaatt cctgcagccc gggggatcca
4741 ctagttctag agcggcccgc ggtaccgtcg acggtaccgc gggcccggga tccaccggat
4801 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
4861 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
4921 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
4981 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
5041 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
5101 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
5161 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
5221 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
5281 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
5341 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
5401 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
5461 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
5521 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
5581 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
5641 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
5701 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
5761 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
5821 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
5881 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
5941 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
6001 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
6061 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
6121 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
6181 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
6241 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
6301 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
6361 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
6421 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
6481 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
6541 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
6601 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
6661 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
6721 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
6781 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
6841 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
6901 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
6961 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
7021 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
7081 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
7141 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
7201 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
7261 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
7321 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
7381 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
7441 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
7501 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
7561 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
7621 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
7681 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
7741 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
7801 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
7861 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
7921 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
7981 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
8041 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
8101 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
8161 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
8221 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
8281 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
8341 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
8401 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
8461 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
8521 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
8581 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
8641 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
8701 ttagtgaacc gtcagatccg ctagcgctac cggtcgccac c
//
