| 质粒名称: | miRFP670nano-pcDNA |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 127443 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA |
| 完整质粒大小: | 5491bp |
| 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: | miRFP670nano |
| 物种来源: | Synthetic |
| 插入大小: | 441 bp |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | KpnI(未破坏) |
| 3’克隆位点: | EcoRI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | miRFP670nano-pcDNA was a gift from Vladislav Verkhusha (Addgene plasmid # 127443 ; http://n2t.net/addgene:127443 ; RRID:Addgene_127443) |
| 参考文献部分: | Smallest near-infrared fluorescent protein evolved from cyanobacteriochrome as versatile tag for spectral multiplexing. Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV. Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8. 10.1038/s41467-018-08050-8 PubMed 30655515 |
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 miRFP670nano-pcDNA 5491 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Monomeric near-infrared fluorescent protein miRFP670nano in
mammalian plasmid.
ACCESSION .
VERSION .
KEYWORDS miRFP670nano-pcDNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5491)
AUTHORS Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
TITLE Smallest near-infrared fluorescent protein evolved from
cyanobacteriochrome as versatile tag for spectral multiplexing.
JOURNAL Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
PUBMED 30655515
REFERENCE 2 (bases 1 to 5491)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5491
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 152..531
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 532..735
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 686..706
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 780..799
/label=T7
/note="T7 promoter, forward primer"
promoter 780..798
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1331..1360
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1376..1393
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1416..1433)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1422..1646
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1692..2120
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1779..1798)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1989..2010
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2129..2149)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2134..2463
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2314..2449
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2376..2395
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 2511..2558
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 2577..2951
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 3081..3202
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3118..3137)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3172..3191
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3251..3267)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3251..3267)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3264..3286)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3275..3291
/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(3299..3329)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3344..3365
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3482..3499)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3653..4241)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3733..3752)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4412..5272)
/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 5035..5054
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5273..5377)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5452..5471)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
61 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
121 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
541 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
661 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
721 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
781 aatacgactc actataggga gacccaagct ggctagttaa gcttggtacc atggcaaacc
841 tggacaagat gctgaatacc acagtaacag aggtgcggca gttcctgcag gtggacagag
901 tgtgcgtgtt ccagtttgag gaggattata gcggagtggt ggtggtggag gccgtggacg
961 ataggtggat ctccatcctg aagacccagg tgcgggatag atacttcatg gagacaaggg
1021 gcgaggagta ttctcacggc cgctaccagg ccatcgccga catctacacc gcaaacctga
1081 cagagtgcta cagggatctg ctgacacagt ttcaggtgag agcaatcctg gccgtgccca
1141 tcctgcaggg caagaagctg tggggcctgt tggtggcaca ccagctggcg gcccctagac
1201 agtggcagac ctgggagatc gactttctga agcagcaggc cgtggtggtg ggcatcgcca
1261 tccagcagag ctagctgaat tctgcagata tccagcacag tggcggccgc tcgagtctag
1321 agggcccttc gaacaaaaac tcatctcaga agaggatctg aatatgcata ccggtcatca
1381 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
1441 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
1501 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
1561 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
1621 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
1681 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
1741 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
1801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
1861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
1921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
1981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
2041 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
2101 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
2161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
2221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
2281 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
2341 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
2401 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
2461 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
2521 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
2581 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
2641 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
2701 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
2761 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
2821 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
2881 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
2941 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
3001 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
3061 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
3121 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
3181 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
3241 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
3301 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
3361 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
3421 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
3481 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
3541 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
3601 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
3661 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
3721 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
3781 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
3841 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
3901 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
3961 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
4021 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
4081 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
4141 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
4201 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
4261 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
4321 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
4381 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
4441 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
4501 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
4561 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
4621 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
4681 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
4741 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
4801 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
4861 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
4921 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
4981 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
5041 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
5101 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
5161 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
5221 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
5281 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
5341 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
5401 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
5461 ctctgatgcc gcatagttaa gccagtatct g
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pmiRFP720-N1
- 下一篇：IFP2.0-N1
