| 质粒名称: | pPSU10-30-50-100 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 173838 |
| 存储实验室: | Song Tan |
|
载体骨架基本信息 |
|
| 载体名称: | pST44 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 8069 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | STRHSTPAB |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | T7 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NdeI(未破坏) |
| 3’克隆位点: | BsrGI (未破坏) |
| 5’测序引物 | T7 |
| 3’测序引物 | T7 Term |
|
文献引用方法 |
|
| 材料和方法部分: | pPSU10-30-50-100 was a gift from Song Tan (Addgene plasmid # 173838 ; http://n2t.net/addgene:173838 ; RRID:Addgene_173838) |
| 参考文献部分: | The Penn State Protein Ladder system for inexpensive protein molecular weight markers. Santilli RT, Williamson JE 3rd, Shibata Y, Sowers RP, Fleischman AN, Tan S. Sci Rep. 2021 Aug 18;11(1):16703. doi: 10.1038/s41598-021-96051-x. 10.1038/s41598-021-96051-x PubMed 34408191 |
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 pPSU10-30-50-100 8069 bp ds-DNA circular SYN 17-AUG-2021
DEFINITION Coexpresses 10, 30, 50 and 100 kD Penn State ladder proteins.
ACCESSION .
VERSION .
KEYWORDS pPSU10-30-50-100
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8069)
AUTHORS Tan, S
TITLE TanLab Protein Ladder Plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 8069)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 17, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8069
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 27..46
/label=T7
/note="T7 promoter, forward primer"
promoter 27..45
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 77..99
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 116..139
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 149..178
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 377..1474
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDEALKDAQT"
regulatory 1331..1340
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1448..1471
/label=MBP-F
/note="Maltose binding protein, forward primer"
primer_bind 1499..1516
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1500..1516
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
RBS 1529..1551
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1568..1591
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 1601..1630
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
RBS 4242..4264
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 4281..4310
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 4995..5072
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
RBS 5122..5144
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5161..5184
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 5194..5223
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
primer_bind complement(5499..5517)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5513..5560
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 5627..5718
/gene="bla"
/label=AmpR promoter
terminator 6583..6622
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin 6653..7241
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7142..7161
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7395..7412
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 7427..7566
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(7492..7511)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7652..7674
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(7668..7859)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 ggcaccatgc atcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
61 tccctctaga aataattttg tttaacttta agaaggagat atacatatgg caagctggtc
121 acatccgcag tttgaaaaag gtagcagtca tcatcaccac catcaccatc atcatcacag
181 cagcggtagc ggtagcgcag ataacaaatt taacaaagaa cagcagaacg ccttctacga
241 aattctgcat ctgccgaatc tgaatgaaga acagcgtaat ggttttatcc agagcctgaa
301 agatgatccg agccagagcg caaatctgct ggccgaagca aaaaaactga atgatgcaca
361 ggcaccgaaa ggatctaaaa tcgaagaagg taaactggta atctggatta acggcgataa
421 aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt
481 caccgttgag cacccggata aactggaaga gaaattccca caggttgcgg caactggcga
541 tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct
601 gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga
661 tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct
721 gatttataac aaggatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct
781 ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta
841 cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa
901 gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct
961 ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc
1021 tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat
1081 cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc
1141 caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct
1201 ggcgaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa
1261 agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc
1321 acgtattgcc gccaccatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca
1381 gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca
1441 gactgtcgat gaagccctga aagacgcgca gactaactaa tgtacaagat ccagatcttg
1501 taaaacgacg gccagtgaat tcaataattt tgtttaactt taagaaggag atatacatat
1561 ggcaagctgg tcacatccgc agtttgaaaa aggtagcagt catcatcacc accatcacca
1621 tcatcatcac agcagcggta gcggtagcgc agataacaaa tttaacaaag aacagcagaa
1681 cgccttctac gaaattctgc atctgccgaa tctgaatgaa gaacagcgta atggttttat
1741 ccagagcctg aaagatgatc cgagccagag cgcaaatctg ctggccgaag caaaaaaact
1801 gaatgatgca caggcaccga aaggatctgc tgataacaaa tttaacaaag agcaacaaaa
1861 cgccttttac gagatcctgc atttaccgaa cctgaccgag gaacagcgca acggctttat
1921 tcagtcactg aaagacgacc cgagtgtgag caaagaaatt ctggcagagg ccaaaaagct
1981 gaacgatgcc caagctccga aaggatccgg tagcagcggt agcagtggtt caagcgcacc
2041 ggttcgtagc ctgaattgta ccctgcgtga tagccagcag aaaagcctgg ttatgagtgg
2101 tccgtatgaa ctgaaagcac tgcatctgca aggtcaggat atggaacagc aggttgtttt
2161 tagcatgagc tttgttcagg gtgaagaaag caatgataaa attccggttg cactgggtct
2221 gaaagagaaa aatctgtatc tgagctgcgt gctgaaagat gataaaccga ccttacagct
2281 ggaaagcgtt gatccgaaaa actacccgaa aaagaaaatg gaaaaacgct ttgtgttcaa
2341 caaaatcgag atcaacaaca aacttgagtt tgagagcgca cagtttccga attggtatat
2401 tagcaccagc caggccgaaa atatgccggt ttttttaggt ggcaccaaag gtggccagga
2461 tattaccgat tttaccatgc agtttgtgtc cagcgccggc gaaggtcagg gccagggtca
2521 aggtgaaggt tccggcagtg aggcagaagc ccgcccgact aactttatcc gtcagatcat
2581 cgatgaagat ctggccagtg gtaagcacac cacagtacac acccgtttcc cgccggagcc
2641 gaatggctat ctgcatattg gccatgcgaa atctatctgc ctgaacttcg ggatcgccca
2701 ggactataaa ggccagtgca acctgcgttt cgacgacact aacccggtaa aagaagatat
2761 cgagtatgtt gagtcgatca aaaacgacgt agagtggtta ggttttcact ggtctggtaa
2821 cgtccgttac tcctccgatt attttgatca gctccacgcc tatgcgatcg aactgatcaa
2881 taaaggcctg gcgtacgttg atgaactgac gccggaacag atccgcgaat accgcggcac
2941 cctgacgcaa ccgggtaaaa acagcccgta ccgcgaccgc agcgttgaag agaacctggc
3001 gctgttcgaa aaaatgcgtg ccggtggttt tgaagaaggt aaagcctgcc tgcgtgcgaa
3061 aatcgacatg gcttcaccgt ttatcgtgat gcgcgatccg gtgctgtacc gtattaaatt
3121 tgctgaacac caccagactg gcaacaagtg gtgcatctac ccgatgtacg acttcaccca
3181 ctgcatcagc gatgccctgg aaggtattac gcactctctg tgtacgcttg agttccagga
3241 caaccgtcgt ctgtacgact gggtactgga caacatcacg attcctgttc acccgcgcca
3301 gtatgagttc tcgcgcctga atctggaata caccgtgatg tccaagcgta agttgaacct
3361 gctggtgacc gacaagcacg ttgaaggctg ggatgacccg cgtatgccga ccatttccgg
3421 tctgcgtcgt cgtggttaca ctgcggcttc tattcgtgag ttctgcaaac gcatcggcgt
3481 gaccaagcag gacaacacca ttgagatggc gtcgctggaa tcctgcatcc gtgaagatct
3541 caacgaaaat gcgccgcgcg caatggcggt tatcgatccg gtgaaactgg ttatcgaaaa
3601 ctatcagggc gaaggcgaaa tggttaccat gccgaaccat ccgaacaaac cggaaatggg
3661 cagccgtcag gtgccgttta gcggtgagat ttggattgat cgcgccgatt tccgcgaaga
3721 agctaacaag cagtacaaac gtctggtgct gggtaaagaa gtgcgtctgc gtaatgctta
3781 tgtgattaag gcagaacgtg tcgagaaaga tgccgaaggt aatatcacca ccatcttctg
3841 tacttatgac gccgatacct taagcaaaga tccggcagat ggtcgtaaag tcaaaggtgt
3901 tattcactgg gtgagcgcgg cacatgcgct gccggttgaa atccgtttgt atgaccgtct
3961 gttcagcgtg cctaacccag gtgctgcgga tgatttcctg tcggtgatta acccggaatc
4021 gctggtgatc aaacagggct ttgctgaacc gtcgctgaaa gatgcggttg cgggtaaagc
4081 attccagttt gagcgtgaag gttacttctg cctcgatagc cgccattcta cggcggaaaa
4141 accggtattt aaccgcaccg ttgggctgcg tgatacctgg gcgaaagtag gcgagtaatg
4201 tacaaagctt cgccagggtt ttcccagtcg agctcaataa ttttgtttaa ctttaagaag
4261 gagatataca tatgggtagt catcatcacc accatcacca tcatcatcat ggtagcgcag
4321 ataacaagtt caataaagaa cagcagaacg ccttctatga aattctgcat ctgccgaatc
4381 tgaatgaaga acagcgtaat ggttttatcc agagcctgaa agatgatccg agccagagcg
4441 caaatctgct ggccgaagca aaaaaactga atgatgcaca ggcaccgaaa ggatccgctg
4501 ataacaaatt taacaaagag caacaaaacg ccttttacga gatcctgcat ttaccgaacc
4561 tgaccgagga acagcgcaac ggctttattc agtcactgaa agacgacccg agtgtgagca
4621 aagaaattct ggcagaggcc aaaaagctga acgatgccca agctccggcc ggaggtagcg
4681 gtagcggttc aggttctggt tcaggtagtg gtagcagcga actggaaaaa gcaatggttg
4741 cactgattga tgtgtttcat cagtatagcg gtcgtgaagg cgataaacac aaactgaaaa
4801 aaagcgagct gaaagagctg atcaataacg aactgagcca ttttctggaa gagatcaaag
4861 aacaagaggt ggtggataaa gttatggaaa ccctggataa tgatggtgat ggtgaatgcg
4921 attttcaaga gtttatggca tttgttgcaa tggttaccac cgcctgtcat gaattttttg
4981 aacacgaagc cggcaagcga cgatggaaaa agaatttcat agccgtctca gcagccaacc
5041 gctttaagaa aatctcatcc tccggggcac ttctgtaatg tacaggtacc agcggataac
5101 aatttcacat ccggaaataa ttttgtttaa ctttaagaag gagatataca tatggcaagc
5161 tggtcacatc cgcagtttga aaaaggtagc agtcatcatc accaccatca ccatcatcat
5221 cacagcagcg gtagcggtag cgcagataac aaatttaaca aagaacagca gaacgccttc
5281 tacgaaattc tgcatctgcc gaatctgaat gaagaacagc gtaatggttt tatccagagc
5341 ctgaaagatg atccgagcca gagcgcaaat ctgctggccg aagcaaaaaa actgaatgat
5401 gcacaggcac cgaaaggatc ctaagccggc tgtacaacgc gtctcgagag atccggctgc
5461 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
5521 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
5581 cgactagtag cccgcctaat gagcgggctt ttttttaatt cccctatttg tttatttttc
5641 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
5701 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
5761 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tcgtgaaagt aaaagacgca
5821 gaggaccaat tgggggcacg agtgggatac atagaactgg acttgaatag cggtaaaatc
5881 cttgagagtt ttcgccctga agagcgtttt ccaatgatga gcactttcaa agttctgcta
5941 tgtggagcag tattatcccg tgtagatgcg gggcaagagc aactcggacg acgaatacac
6001 tattcgcaga atgacttggt tgaatactcc ccagtgacag aaaagcacct tacggacgga
6061 atgacggtaa gagaattatg tagtgccgcc ataacgatga gtgataacac tgcggcgaac
6121 ttacttctga caaccatcgg tggaccgaag gaattaaccg cttttttgca caatatggga
6181 gaccatgtaa ctcgccttga ccgttgggaa ccagaactga atgaagccat accaaacgac
6241 gagcgagaca ccacaatgcc tgcggcaatg gcaacaacat tacgcaaact attaactggc
6301 gaactactta ctctggcttc acggcaacaa ttaatagact ggcttgaagc ggataaagtt
6361 gcaggaccac tactgcgttc ggcacttcct gctggctggt ttattgctga taaatctggg
6421 gcaggagagc gtggttcacg gggtatcatt gccgcacttg gaccagatgg taagccttcc
6481 cgtatcgtag ttatctacac gacgggtagt caggcaacta tggacgaacg aaatagacag
6541 attgctgaaa taggggcttc actgattaag cattggtaaa ccgatacaat taaaggctcc
6601 ttttggagcc tttttttttg gacggaccgg tagaaaagat caaaggatct tcttgagatc
6661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
6721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
6781 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
6841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
6901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
6961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
7021 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
7081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
7141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
7201 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
7261 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
7321 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
7381 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
7441 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
7501 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
7561 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
7621 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
7681 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
7741 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
7801 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
7861 gatgcctccg tgtaaggggg atttctgttc atggggtaat gataccgatg aaacgagaga
7921 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
7981 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
8041 agcgcttcgt taatacagat gtaggtgtt
//
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