| 质粒名称: | pCS-H2B-mRFP1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 53745 |
| 存储实验室: | Sean Megason |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+ |
| 载体出品公司: | RZPD |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4095 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | histone H2B,histone cluster 2, H2be |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | simian CMV IE94 |
| 标签/融合蛋白: | mRFP（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | SP6 |
| 3' 测序引物 | EBV-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pCS-H2B-mRFP1 was a gift from Sean Megason (Addgene plasmid # 53745 ; http://n2t.net/addgene:53745 ; RRID:Addgene_53745) |
| 参考文献部分: | In toto imaging of embryogenesis with confocal time-lapse microscopy. Megason SG. Methods Mol Biol. 2009;546:317-32. doi: 10.1007/978-1-60327-977-2_19. 10.1007/978-1-60327-977-2_19 PubMed 19378112 |
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 pCS-H2B-mRFP1 5186 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Contains a fusion of human histone H2B to monomeric red fluorescent
protein 1 in the pCS2+ expression vector.
ACCESSION .
VERSION .
KEYWORDS pCS-H2B-mRFP1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5186)
AUTHORS Megason SG
TITLE In toto imaging of embryogenesis with confocal time-lapse
microscopy.
JOURNAL Methods Mol Biol. 2009;546:317-32. doi:
10.1007/978-1-60327-977-2_19.
PUBMED 19378112
REFERENCE 2 (bases 1 to 5186)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5186
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 57..1034
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1069..1087
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1069..1086
/label=SP6
/note="SP6 promoter, forward primer"
promoter complement(1156..1173)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1192..1569
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAITKYTSAK"
regulatory 1582..1591
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1588..2265
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(1718..1736)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1986..2005
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2180..2203
/label=DsRed1-C
/note="DsRed1, forward primer"
polyA_signal 2272..2406
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2296..2315)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2350..2369
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2519..2539)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2519..2537)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2558..2574)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2558..2574)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2571..2593)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2582..2598
/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(2606..2636)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2651..2672
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2789..2806)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2960..3548)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3040..3059)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3719..4579)
/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 4342..4361
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4580..4684)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4710..5165)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4847..4868)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5059..5078
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca
61 attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca
121 attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca
181 attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca
241 actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc
301 aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca
361 ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt
421 caatactatg tattggccct atgccatata gtattccata tatgggtttt cctattgacg
481 tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc
541 catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca
601 tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg
661 gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg
721 atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa
781 tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt
841 ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc
901 gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg
961 gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg
1021 ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat ttaggtgaca
1081 ctatagaata caagctactt gttctttttg caggatccca tcgattcgaa ttcaaggcct
1141 ctcgagcctc tagaactata gtgagtcgta ttactcgacg gtaccgccac catgccagag
1201 ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac taaggcgcag
1261 aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga gctattccat ctatgtgtac
1321 aaggttctga agcaggtcca ccctgacacc ggcatttcgt ccaaggccat gggcatcatg
1381 aattcgtttg tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat
1441 tacaacaagc gctcgaccat cacctccagg gagatccaga cggccgtgcg cctgctgctg
1501 cctggggagt tggccaagca cgccgtgtcc gagggtacta aggccatcac caagtacacc
1561 agcgctaagg atccaccggt cgccaccatg gcctcctccg aggacgtcat caaggagttc
1621 atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
1681 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
1741 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg ctccaaggcc
1801 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
1861 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
1921 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
1981 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcggatgtac
2041 cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa ggacggcggc
2101 cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc
2161 gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg
2221 gaacagtacg agcgcgccga gggccgccac tccaccggcg cctaatacgt agatccagac
2281 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
2341 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
2401 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
2461 gttttttaat tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc
2521 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
2581 attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct
2641 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
2701 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
2761 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
2821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
2881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
2941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
3001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
3061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
3121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
3181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
3241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
3301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
3361 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
3421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
3481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
3541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
3601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
3661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
3721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
3781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
3841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
3901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
3961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
4021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
4081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
4141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
4201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
4261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
4321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
4381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
4441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
4501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
4561 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
4621 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
4681 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
4741 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
4801 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
4861 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
4921 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
4981 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
5041 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
5101 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
5161 cgcgtcccat tcgccattca ggctgc
//
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