| 质粒名称: | pYD1-mSA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 39865 |
| 存储实验室: | Sheldon Park |
|
载体骨架基本信息 |
|
| 载体名称: | pYD1 |
| 载体出品公司: | invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达 |
| 筛选标记: | TRP1 |
| 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: | monomeric streptavidin,streptavidin rhizavidin hybrid |
| 物种来源: | Synthetic |
| 插入大小: | 357 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
Aga2（主干上的N终端）
HA（插入件上的N端子）
FLAG（主干上的C终端
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pYD1-mSA was a gift from Sheldon Park (Addgene plasmid # 39865 ; http://n2t.net/addgene:39865 ; RRID:Addgene_39865) |
| 参考文献部分: | Stable, high-affinity streptavidin monomer for protein labeling and monovalent biotin detection. Lim KH, Huang H, Pralle A, Park S. Biotechnol Bioeng. 2013 Jan;110(1):57-67. doi: 10.1002/bit.24605. Epub 2012 Aug 8. 10.1002/bit.24605 PubMed 22806584 |
买家导航
pYD1mSA载体基本信息
pYD1mSA载体图谱质粒图谱和多克隆位点信息
pYD1mSA载体简介
pYD1mSA载体序列
LOCUS pYD1-mSA 6490 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pYD1-mSA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6490)
AUTHORS Lim KH, Huang H, Pralle A, Park S
TITLE Stable, high-affinity streptavidin monomer for protein labeling and
monovalent biotin detection.
JOURNAL Biotechnol Bioeng. 2013 Jan;110(1):57-67. doi: 10.1002/bit.24605.
Epub 2012 Aug 8.
PUBMED 22806584
REFERENCE 2 (bases 1 to 6490)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6490
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 138..149
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 150..176
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 240..581
/codon_start=1
/product="monomeric streptavidin/rhizavidin hybrid (Lim et
al., 2013)"
/label=mSA
/translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
QGQDTFTKVK"
CDS 603..626
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(956..975)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(957..975)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(982..999)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(982..998)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(991..1013)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 1139..1594
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1226..1245)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1436..1457
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(1697..2371)
/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(2372..2652)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
primer_bind complement(2673..2692)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2792..2814
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2852..2870)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 2911..3414
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 3451..3555
/gene="bla"
/label=AmpR promoter
CDS 3556..4416
/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 complement(3774..3793)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4587..5175
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5076..5095
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5329..5346
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5463..5484
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5499..5529
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5537..5553
/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)."
primer_bind 5542..5564
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5561..5577
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5561..5577
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 5598..5616
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 5646..6310
/gene="GAL1 and GAL10 genes of S. cerevisiae"
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
protein_bind 5862..5979
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 6274..6297
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
1 acgggaaggc aatgcaagga gtttttgaat attacaaatc agtaacgttt gtcagtaatt
61 gcggttctca cccctcaaca actagcaaag gcagccccat aaacacacag tatgttttta
121 aggacaatag ctcgacgatt gaaggtagat acccatacga cgttccagac tacgctctgc
181 aggctagtgg tggtggtggt tctggtggtg gtggttctgg tggtggtggt tctgctagcg
241 cggaagcggg tatcaccggc acgtggtaca accagtctgg ttctaccttc accgttaccg
301 cgggtgcgga cggtaacctg accggtcagt acgaaaaccg tgcgcagggc actggttgcc
361 agaactctcc gtacaccctg accggtcgtt acaacggtac caaactggaa tggcgtgttg
421 aatggaacaa ctctaccgaa aactgccact ctcgtaccga atggcgtggt cagtaccagg
481 gtggtgcgga agcgcgtatc aacacccagt ggaacctgac ctacgaaggt ggttctggtc
541 cggcgaccga acagggtcag gacaccttca ccaaagttaa accgtctgcg gcgtctggat
601 ccgactacaa ggacgatgac gacaagtaac tcgagatctg ataacaacag tgtagatgta
661 acaaaatcga ctttgttccc actgtacttt tagctcgtac aaaatacaat atacttttca
721 tttctccgta aacaacatgt tttcccatgt aatatccttt tctatttttc gttccgttac
781 caactttaca catactttat atagctattc acttctatac actaaaaaac taagacaatt
841 ttaattttgc tgcctgccat atttcaattt gttataaatt cctataattt atcctattag
901 tagctaaaaa aagatgaatg tgaatcgaat cctaagagaa ttgagctcca attcgcccta
961 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
1021 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
1081 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
1141 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
1201 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
1261 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
1321 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
1381 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
1441 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
1501 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
1561 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
1621 atctgtgcgg tatttcacac cgcatagatc ggcaagtgca caaacaatac ttaaataaat
1681 actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt
1741 cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc
1801 taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagcttt
1861 cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg
1921 tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac
1981 tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga
2041 actcttggta ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat
2101 cattgaccag agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt
2161 tcggagtgcc tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa
2221 ccgggtcaat tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg
2281 aatctagagc acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac
2341 cagaactacc tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca
2401 cgtatactca cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt
2461 tttcgaccga attaattctt aatcggcaaa aaaagaaaag ctccggatca agattgtacg
2521 taaggtgaca agctattttt caataaagaa tatcttccac tactgccatc tggcgtcata
2581 actgcaaagt acacatatat tacgatgctg ttctattaaa tgcttcctat attatatata
2641 tagtaatgtc gtgatctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
2701 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
2761 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
2821 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
2881 aatgtcatga taataatggt ttcttagacg gatcgcttgc ctgtaactta cacgcgcctc
2941 gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt
3001 tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa
3061 aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag
3121 acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca
3181 caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct
3241 gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta
3301 catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa
3361 tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag
3421 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
3481 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
3541 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
3601 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
3661 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
3721 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
3781 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
3841 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
3901 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
3961 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
4021 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
4081 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
4141 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
4201 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
4261 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
4321 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
4381 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
4441 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
4501 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
4561 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
4621 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
4681 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
4741 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
4801 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
4861 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
4921 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
4981 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
5041 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
5101 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
5161 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
5221 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
5281 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
5341 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
5401 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
5461 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
5521 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
5581 gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctggt accaattcct
5641 tgaattttca aaaattctta cttttttttt ggatggacgc aaagaagttt aataatcata
5701 ttacatggca ttaccaccat atacatatcc atatacatat ccatatctaa tcttacttat
5761 atgttgtgga aatgtaaaga gccccattat cttagcctaa aaaaaccttc tctttggaac
5821 tttcagtaat acgcttaact gctcattgct atattgaagt acggattaga agccgccgag
5881 cgggtgacag ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg
5941 ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata
6001 ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa
6061 atgaacgaat caaattaaca accataggat gataatgcga ttagtttttt agccttattt
6121 ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat ataaatgcaa
6181 aaactgcata accactttaa ctaatacttt caacattttc ggtttgtatt acttcttatt
6241 caaatgtaat aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag
6301 gagaaaaacc ccggatcgaa ttccctactt catacatttt caattaagat gcagttactt
6361 cgctgttttt caatattttc tgttattgct tcagttttag cacaggaact gacaactata
6421 tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt actctttgtc aacgactact
6481 attttggcca
//
