| 质粒名称: | pmScarlet_Giantin_C1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 85048 |
| 存储实验室: | Gadella Lab |
|
载体骨架基本信息 |
|
| 载体名称: | pmScarlet_C1 |
| 载体出品公司: | Gadella Lab |
| 空载体大小: | 4671bp |
| 完整质粒大小: | 5061 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Giantin/Golgin B1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 390bp |
| 基因突变: | aa3131-3259 |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BsrGI (未破坏) |
| 3’克隆位点: | BamHI (未破坏) |
| 5’测序引物 | AGGTCTATATAAGCAGAGC |
| 3’测序引物 | TCTACAAATGTGGTATGGC |
|
文献引用方法 |
|
| 材料和方法部分: | pmScarlet_Giantin_C1 was a gift from Dorus Gadella (Addgene plasmid # 85048 ; http://n2t.net/addgene:85048 ; RRID:Addgene_85048) |
| 参考文献部分: | mScarlet: a bright monomeric red fluorescent protein for cellular imaging. Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M, Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr. Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074. 10.1038/nmeth.4074 PubMed 27869816 |
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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 pmScarlet_Giantin_C1 5061 bp ds-DNA circular SYN 23-SEP-2022
DEFINITION In vivo visualization of the Golgi apparatus (can be used for
colocalization studies).
ACCESSION .
VERSION .
KEYWORDS pmScarlet_Giantin_C1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5061)
AUTHORS Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M,
Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr
TITLE mScarlet: a bright monomeric red fluorescent protein for cellular
imaging.
JOURNAL Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074.
PUBMED 27869816
REFERENCE 2 (bases 1 to 5061)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5061
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 61..165
/gene="bla"
/label=AmpR promoter
promoter 167..524
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(190..210)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 375..510
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 437..456
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 559..1353
/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(613..632)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1223..1242
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(1541..1560)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 1585..1632
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 1961..2549
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2450..2469
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 2724..3027
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3028..3231
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3182..3202
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 3270..3279
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3276..3971
/codon_start=1
/product="bright monomeric red fluorescent protein evolved
from a synthetic template (Bindels et al., 2016)"
/label=mScarlet
/translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
STGGMDELYK"
primer_bind complement(3409..3427)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3677..3696
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 4512..4633
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4549..4568)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4603..4622
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(4640..34)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4777..4798)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4989..5008
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
61 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
181 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
241 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
301 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
361 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
421 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
481 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
541 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
601 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
661 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
721 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
781 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
841 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
901 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
961 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
1021 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
1081 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
1141 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
1201 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
1261 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
1321 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
1381 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
1441 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
1501 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
1561 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
1621 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
1681 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
1741 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
1801 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
1861 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
1921 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
1981 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
2041 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
2101 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
2161 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
2221 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
2281 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
2341 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
2401 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
2461 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
2521 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
2581 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
2641 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
2701 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
2761 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
2821 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
2881 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
2941 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
3001 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
3061 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
3121 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
3181 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
3241 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggcagtga
3301 tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg
3361 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
3421 tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg
3481 gctccagggc cttcaccaag caccccgccg acatccccga ctactataag cagtccttcc
3541 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga
3601 cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca
3661 acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg
3721 agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga
3781 aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc
3841 agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact
3901 acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg
3961 agctgtacaa gtccggactc agatctcgag gagaaccgca gcaaagcttt tctgaagctc
4021 agcagcagct atgcaacacc agacaggaag tgaatgaatt aaggaagctg ctggaagaag
4081 aacgagacca aagagtggct gctgagaatg ctctctctgt ggccgaggag cagatcagac
4141 ggttagagca cagtgaatgg gactcttccc ggactcctat cattggctcc tgtggcactc
4201 aggagcaggc actgttaata gatcttacaa gcaacagttg tcgaaggacc cggagtggcg
4261 ttggatggaa gcgagtcctg cgttcactct gtcattcacg gacccgagtg ccacttctag
4321 cagccatcta ctttctaatg attcatgtcc tgctcattct gtgttttacg ggccatctat
4381 agggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
4441 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
4501 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
4561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
4621 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
4681 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
4741 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
4801 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
4861 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
4921 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
4981 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
5041 acgctgcgcg taaccaccac a
//
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