| 质粒名称: | YPet-N1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 54637 |
| 存储实验室: | Patrick Daugherty, Michael Davidson |
|
载体骨架基本信息 |
|
| 载体名称: | YPet-N1 |
| 空载体大小: | 4750 bp |
| 完整质粒大小: | 4733 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: | NO |
| 标签/融合蛋白: | YPet (C terminal on backbone) |
|
文献引用方法 |
|
| 材料和方法部分: | YPet-N1 was a gift from Patrick Daugherty & Michael Davidson (Addgene plasmid # 54637 ; http://n2t.net/addgene:54637 ; RRID:Addgene_54637) |
| 参考文献部分: | Evolutionary optimization of fluorescent proteins for intracellular FRET. Nguyen AW, Daugherty PS. Nat Biotechnol. 2005 Mar . 23(3):355-60. 10.1038/nbt1066 PubMed 15696158 |
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 YPet-N1 4733 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Localization: N1 Cloning Vector, Excitation: 513, Emission: 527.
ACCESSION .
VERSION .
KEYWORDS YPet-N1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4733)
AUTHORS Nguyen AW, Daugherty PS
TITLE Evolutionary optimization of fluorescent proteins for intracellular
FRET.
JOURNAL Nat Biotechnol. 2005 Mar . 23(3):355-60.
PUBMED 15696158
REFERENCE 2 (bases 1 to 4733)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4733
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 598..678
/label=MCS
/note="multiple cloning site"
regulatory 680..689
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 686..1405
/codon_start=1
/product="yellow fluorescent protein optimized for FRET
with CyPet (Nguyen and Daugherty, 2005)"
/label=YPet
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
EFLTAAGITEGMNELYK"
primer_bind complement(992..1011)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
polyA_signal 1528..1649
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1565..1584)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1619..1638
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1656..2111)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1793..1814)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2005..2024
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2138..2242
/gene="bla"
/label=AmpR promoter
promoter 2244..2601
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2267..2287)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2452..2587
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2514..2533
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2636..3430
/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(2690..2709)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3300..3319
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3618..3637)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3662..3709
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4038..4626
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4527..4546
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
661 ggcccgggat ccaccggtcg ccaccatggt gagcaaaggc gaagagctgt tcaccggcgt
721 ggtgcccatc ctggtggagc tggacggcga cgtgaacggc cacaagttca gcgtgagcgg
781 cgagggcgag ggcgacgcca cctacggcaa gctgaccctg aagctgctgt gcaccaccgg
841 caagctgccc gtgccctggc ccaccctggt gaccaccctg ggctacggcg tgcagtgctt
901 cgcccggtac cccgaccaca tgaagcagca cgacttcttc aagagcgcca tgcccgaggg
961 ctacgtgcag gagcggacca tcttcttcaa ggacgacggc aactacaaga cccgggccga
1021 ggtgaagttc gagggcgaca ccctggtgaa ccggatcgag ctgaagggca tcgacttcaa
1081 ggaggacggc aacatcctgg gccacaagct ggagtacaac tacaacagcc acaacgtgta
1141 catcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc ggcacaacat
1201 cgaggacggc ggcgtgcagc tggccgacca ctaccagcag aacaccccca tcggcgacgg
1261 ccccgtgctg ctgcccgaca accactacct gagctaccag agcgccctgt tcaaggaccc
1321 caacgagaag cgggaccaca tggtgctgct ggagttcctg accgccgccg gcatcaccga
1381 gggcatgaac gagctctata agtaaagcgg ccgcgactct agatcataat cagccatacc
1441 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
1501 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
1561 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
1621 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
1681 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
1741 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
1801 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
1861 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
1921 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
1981 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
2041 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
2101 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
2161 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
2221 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
2281 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
2341 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
2401 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
2461 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
2521 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
2581 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
2641 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
2701 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
2761 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
2821 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
2881 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
2941 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
3001 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
3061 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
3121 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
3181 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
3241 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
3301 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
3361 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
3421 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
3481 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
3541 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
3601 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
3661 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
3721 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
3781 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
3841 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
3901 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
3961 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
4021 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
4081 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
4141 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
4201 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
4261 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
4321 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
4381 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
4441 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
4501 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
4561 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
4621 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
4681 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgc
//
