| 质粒名称: | pJYDN3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 162456 |
| 存储实验室: | Gideon Schreiber |
|
载体骨架基本信息 |
|
| 载体名称: | New backbone pJYD |
| 载体出品公司: | Dr. Jiri Zahradnik |
| 空载体大小: | 5254 bp |
| 完整质粒大小: | 5255bp |
| 载体类型: | Shuttle vector bacteria/yeast |
| 筛选标记: | TRP1 |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 标签/融合蛋白: |
ALFA tag (N terminal on backbone)
HA, myc tags (C terminal on backbone)
eUnaG2 fluorescent protein (C terminal on backbone)
|
|
文献引用方法 |
|
| 材料和方法部分: | pJYDN3 was a gift from Gideon Schreiber (Addgene plasmid # 162456 ; http://n2t.net/addgene:162456 ; RRID:Addgene_162456) |
| 参考文献部分: | An enhanced yeast display platform demonstrates the binding plasticity under various selection pressures. Zahradník J, Dey D, Marciano S, Schreiber G. bioRxiv Preprint 2020 10.1101/2020.12.16.423176 |
买家导航
pJYDN3载体质粒基本信息
pJYDN3质粒图谱载体图谱和pJYDN3载体序列质粒序列多克隆位点信息
pJYDN3质粒载体简介
pJYDN3质粒序列载体序列
LOCUS pJYDN3 5255 bp DNA circular SYN 14-APR-2025
DEFINITION Yeast display platform plasmid N3 which allows for simultaneous
expression of the protein of interest at the N-terminus of Aga2p
yeast agglutinin and eUnaG2 or nbALFA detection..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5255)
AUTHORS Zahradnik J, Dey D, Marciano S, Schreiber G
TITLE An enhanced yeast display platform demonstrates the binding
plasticity under various selection pressures
JOURNAL bioRxiv Preprint 2020
REFERENCE 2 (bases 1 to 5255)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv
Preprint 2020"
COMMENT Sequence Label: pJYDN3
FEATURES Location/Qualifiers
source 1..5255
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(199..873)
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
promoter complement(874..1154)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
primer_bind complement(1175..1194)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(1255..1273)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 1314..1818
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 1855..1959
/gene="bla"
/label=AmpR promoter
CDS 1960..2775
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS 1960..2775
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(2033..2052)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin 2946..3534
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3435..3454
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 3717..4158
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 3717..3834
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 4129..4152
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
CDS 4280..4318
/codon_start=1
/product="rationally designed epitope tag that forms a
stable alpha-helix (Gotzke et al., 2019)"
/label=ALFA-tag
/note="It may be beneficial to flank the ALFA-tag sequence
with proline residues to preserve the helical structure."
/translation="SRLEEELRRRLTE"
CDS 4660..4671
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 4672..4698
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 4765..4794
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
ORIGIN
1 tttgttccca ctgtactttt agctcgtaca aaatacaata tacttttcat ttctccgtaa
61 acaacatgtt ttcccatgta atatcctttt ctatttttcg ttccgttacc aactttacac
121 atacttttgc ggtatttcac accgcataga tcggcaagtg cacaaacaat acttaaataa
181 atactactca gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga
241 gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa
301 tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagct
361 ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca aaagttcacc
421 tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct gtgaagctgc
481 actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg gcaaaccgag
541 gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat caatgccgta
601 atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc caaccaagta
661 tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt tccttgcaat
721 aaccgggtca attgttctct ttctattggg cacacatata atacccagca agtcagcatc
781 ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg
841 accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg tgtgcttaat
901 cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct ctccttttct
961 tttttcgacc gaattaattc ttaatcggca aaaaaagaaa agctccggat caagattgta
1021 cgtaaggtga caagctattt ttcaataaag aatatcttcc actactgcca tctggcgtca
1081 taactgcaaa gtacacatat attacgatgc tgttctatta aatgcttcct atattatata
1141 tatagtaatg tcgtgatcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
1201 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctcgagacga aagggcctcg
1261 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acggatcgct
1321 tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc
1381 tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga
1441 agagttacgg aatgaagaaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc
1501 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt
1561 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga
1621 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt
1681 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt
1741 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt
1801 atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga
1861 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
1921 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagccatat tcaacgggaa
1981 acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
2041 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
2101 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
2161 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
2221 atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc
2281 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
2341 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
2401 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
2461 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
2521 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
2581 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
2641 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
2701 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
2761 gatgagtttt tctaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
2821 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
2881 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
2941 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
3001 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
3061 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
3121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
3181 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
3241 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
3301 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
3361 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
3421 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
3481 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg aaaaaacgcc
3541 agcaacgcgg cactttgtaa ctgagctgtc attttgctgg cccagctggt cgaccgcggc
3601 taagcagtac tggtaccgtt gtggaaatgt aaagagcccc attatcttag cctaaaaaaa
3661 ccttctcttt ggaactttca gtaatacgct taactgctca ttgctatatt gaagtacgga
3721 ttagaagccg ccgagcgggt gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg
3781 tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat
3841 aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca gtaacctggc
3901 cccacaaacc ttcaaatgaa cgaatcaaat taacaaccat aggatgataa tgcgattagt
3961 tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga tctattaaca
4021 gatatataaa tgcaaaaact gcataaccac tttaactaat actttcaaca ttttcggttt
4081 gtattacttc ttattcaaat gtaataaaag tatcaacaaa aaattgttaa tatacctcta
4141 tactttaacg tcaaggagaa aaaaccccgg atcgaattcc ctacttcata cattttcaat
4201 taagatgagg ttcccatcta ttttcaccgc tgttgttttt gctgcttctt ctgctttggc
4261 tgctccagct aatggtccat ccagattgga agaagaattg agaagaagat taaccgaacc
4321 acatatggaa gcataataat taaggcgcct tatctcgagg ggggcggatc ctctggtgga
4381 ggtggttcgg gcggaggtgg taacggatct aatggttctg gcggtagctc tgcacaggaa
4441 ctgacaacta tatgcgagca aatcccctca ccaactttag aatcgacgcc gtactctttg
4501 tcaacgacta ctattttggc caacgggaag gcaatgcaag gagtttttga atattacaaa
4561 tcagtaacgt ttgtcagtaa ttgcggttct cacccctcaa caactagcaa aggcagcccc
4621 ataaacacac agtatgtttt taaggacaat agctcgacga ttgaaggtag atacccatac
4681 gacgttccag actacgctct gcaggctagt ggtggaggag gctctggtgg aggtggtagc
4741 ggaggcggag ggtcggctag ccatgaacag aagttgatct ctgaagaaga tttgatgtta
4801 gaaaaatttg ttggcacctg gaagatcgaa tcctctgaaa attttggtga atacttgaag
4861 gctatcggtg ccccaaaaga attggctgat gctggtgatg ctactactcc agtcttgtac
4921 atttctcaaa aggatggtga taagatgacc gtcaagattg aaaacggtcc accaactttt
4981 ttggataccc aagtttcttt caagttgggt gaagaattcg acgaatttcc atccgataga
5041 agaaagggtg ttaagtccgt tgttaacttg tctggtgaga agttggttta cgttcaaaag
5101 tgggatggta aagaaaccac ttacgtcaga gaaatcaagg acggtaaatt ggttgttact
5161 ttgaccatgg gtgatgttgt tgctgttaga tcttatagaa gggcctctga ataatagaga
5221 tctgataaca acagtgtaga tgtaacaaaa tcgac
//
