| 质粒名称: | pPSU20-40-60-80 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 173839 |
| 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: | 8438 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: | STRHSTPABPACCBPv3 |
| 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(未知是否破坏) |
| 3’克隆位点: | BsrGI (未知是否破坏) |
| 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: | pPSU20-40-60-80 was a gift from Song Tan (Addgene plasmid # 173839 ; http://n2t.net/addgene:173839 ; RRID:Addgene_173839) |
| 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 pPSU20-40-60-80 8438 bp ds-DNA circular SYN 17-AUG-2021
DEFINITION Coexpresses 20, 40, 60 and 80 kD Penn State ladder proteins.
ACCESSION .
VERSION .
KEYWORDS pPSU20-40-60-80
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8438)
AUTHORS Tan, S
TITLE TanLab Protein Ladder Plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 8438)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 17, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8438
/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"
primer_bind 668..685
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 669..685
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
RBS 698..720
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 737..766
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 1130..1783
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
primer_bind 1738..1760
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
RBS 1827..1849
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1866..1889
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 1899..1928
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
RBS 4102..4124
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 4141..4170
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
primer_bind complement(5868..5886)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5882..5929
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 5996..6087
/gene="bla"
/label=AmpR promoter
terminator 6952..6991
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin 7022..7610
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7511..7530
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7764..7781
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 7796..7935
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(7861..7880)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8021..8043
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(8037..8228)
/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 ggatctgctg ataacaaatt taacaaagag caacaaaacg ccttttacga
421 gatcctgcat ttaccgaacc tgaccgagga acagcgcaac ggctttattc agtcactgaa
481 agacgacccg agtgtgagca aagaaattct ggcagaggcc aaaaagctga acgatgccca
541 agctccgaaa ggatccggtg ccggccagga acagctgcaa cagaatttca tagccgtctc
601 agcagccaac cgctttaaga aaatctcatc ctccggggca cttctgtaat gtacaagatc
661 cagatcttgt aaaacgacgg ccagtgaatt caataatttt gtttaacttt aagaaggaga
721 tatacatatg ggtagtcatc atcaccacca tcaccatcat catcatggta gcgcagataa
781 caagttcaat aaagaacagc agaacgcctt ctatgaaatt ctgcatctgc cgaatctgaa
841 tgaagaacag cgtaatggtt ttatccagag cctgaaagat gatccgagcc agagcgcaaa
901 tctgctggcc gaagcaaaaa aactgaatga tgcacaggca ccgaaaggat ccgctgataa
961 caaatttaac aaagagcaac aaaacgcctt ttacgagatc ctgcatttac cgaacctgac
1021 cgaggaacag cgcaacggct ttattcagtc actgaaagac gacccgagtg tgagcaaaga
1081 aattctggca gaggccaaaa agctgaacga tgcccaagct ccggccggct cccctatact
1141 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
1201 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
1261 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
1321 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
1381 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
1441 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
1501 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
1561 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
1621 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
1681 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
1741 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa taatgtacaa agcttcgcca
1801 gggttttccc agtcgagctc aataattttg tttaacttta agaaggagat atacatatgg
1861 caagctggtc acatccgcag tttgaaaaag gtagcagtca tcatcaccac catcaccatc
1921 atcatcacag cagcggtagc ggtagcgcag ataacaaatt taacaaagaa cagcagaacg
1981 ccttctacga aattctgcat ctgccgaatc tgaatgaaga acagcgtaat ggttttatcc
2041 agagcctgaa agatgatccg agccagagcg caaatctgct ggccgaagca aaaaaactga
2101 atgatgcaca ggcaccgaaa ggatctgctg ataacaaatt taacaaagag caacaaaacg
2161 ccttttacga gatcctgcat ttaccgaacc tgaccgagga acagcgcaac ggctttattc
2221 agtcactgaa agacgacccg agtgtgagca aagaaattct ggcagaggcc aaaaagctga
2281 acgatgccca agctccgaaa ggatccggta gcagcggtag cagtggttca agcgcaccgg
2341 ttcgtagcct gaattgtacc ctgcgtgata gccagcagaa aagcctggtt atgagtggtc
2401 cgtatgaact gaaagcactg catctgcaag gtcaggatat ggaacagcag gttgttttta
2461 gcatgagctt tgttcagggt gaagaaagca atgataaaat tccggttgca ctgggtctga
2521 aagagaaaaa tctgtatctg agctgcgtgc tgaaagatga taaaccgacc ttacagctgg
2581 aaagcgttga tccgaaaaac tacccgaaaa agaaaatgga aaaacgcttt gtgttcaaca
2641 aaatcgagat caacaacaaa cttgagtttg agagcgcaca gtttccgaat tggtatatta
2701 gcaccagcca ggccgaaaat atgccggttt ttttaggtgg caccaaaggt ggccaggata
2761 ttaccgattt taccatgcag tttgtgtcca gcgccggcag aaaggccctg actaataaca
2821 acaatgccgg tgaagcggag cagcagccac cgaaggcgaa gcgcgccaga atcgcttttc
2881 atctggagct gcccaagcgg aggaccgtgc tcggcaacgt gctagtctgc ggcaacggcg
2941 atgtgggcca acttggtctc ggcgaggaca tactggagcg caagcgtctc agccctgtgg
3001 cgggcatccc ggatgccgtg gacatatccg ccggtggcat gcacaacctg gtgctgacca
3061 agtcagggga catttactca ttcggttgca atgacgaagg tgctctaggc cgtgacacca
3121 gcgaggatgg cagcgagtca aagccggatc ttatcgattt gcccggcaag gctctgtgca
3181 tttcggcggg agactcgcat tccgcttgtc tgctggagga tggccgtgtc ttcgcttggg
3241 gctcgttccg ggactctcat ggcaacatgg gcctcaccat cgatggtaac aagcgtacgc
3301 ccatcgatct gatggagggc actgtctgtt gtagcatcgc gtccggcgcc gatcatttgg
3361 tcatcctgac cactgccgga aaggtcttta ccgtgggctg cgctgagcag ggacaactcg
3421 ggcggctgtc ggagcgttcc atctccggag agggtcgtcg tggcaaacgc gacttgctgc
3481 ggcccaccca gctcatcatc acgcgggcca aaccttttga ggccatctgg gccaccaact
3541 actgcacatt catgcgcgag tcacagacac aggtcatctg ggccacaggg cttaacaact
3601 tcaagcagct ggcccacgag acaaagggca aggagttcgc actgacgccc atcaagacgg
3661 aactcaaaga tatccgacat attgccgggg gtcagcacca tactgtcatc ctgacaacgg
3721 accttaagtg ttccgtagtg ggtcgtccgg agtacggtcg gcttggtctg ggcgatgtca
3781 aggatgttgt ggagaagcca acgattgtaa agaagctgac cgaaaagatt gtgtccgttg
3841 gatgtggcga agtctgttcc tatgccgtta caattgatgg caaactgtat tcctggggct
3901 ccggtgtcaa caatcaacta ggcgttggcg acggcgatga tgaactcgaa ccgatcgtgg
3961 ttgtcagcaa gaacacgcag gggaagcaca tgctgctcgc atctggcggt ggtcagcacg
4021 caatcttttt ggtcaaagcg gacaaacagg accagtaatg tacaggtacc agcggataac
4081 aatttcacat ccggaaataa ttttgtttaa ctttaagaag gagatataca tatgggtagt
4141 catcatcacc accatcacca tcatcatcat ggtagcgcag ataacaagtt caataaagaa
4201 cagcagaacg ccttctatga aattctgcat ctgccgaatc tgaatgaaga acagcgtaat
4261 ggttttatcc agagcctgaa agatgatccg agccagagcg caaatctgct ggccgaagca
4321 aaaaaactga atgatgcaca ggcaccgaaa ggatccgctg ataacaaatt taacaaagag
4381 caacaaaacg ccttttacga gatcctgcat ttaccgaacc tgaccgagga acagcgcaac
4441 ggctttattc agtcactgaa agacgacccg agtgtgagca aagaaattct ggcagaggcc
4501 aaaaagctga acgatgccca agctccggcc ggaagaaagg ccctgactaa taacaacaat
4561 gccggtgaag cggagcagca gccaccgaag gcgaagcgcg ccagaatcgc ttttcatctg
4621 gagctgccca agcggaggac cgtgctcggc aacgtgctag tctgcggcaa cggcgatgtg
4681 ggccaacttg gtctcggcga ggacatactg gagcgcaagc gtctcagccc tgtggcgggc
4741 atcccggatg ccgtggacat atccgccggt ggcatgcaca acctggtgct gaccaagtca
4801 ggggacattt actcattcgg ttgcaatgac gaaggtgctc taggccgtga caccagcgag
4861 gatggcagcg agtcaaagcc ggatcttatc gatttgcccg gcaaggctct gtgcatttcg
4921 gcgggagact cgcattccgc ttgtctgctg gaggatggcc gtgtcttcgc ttggggctcg
4981 ttccgggact ctcatggcaa catgggcctc accatcgatg gtaacaagcg tacgcccatc
5041 gatctgatgg agggcactgt ctgttgtagc atcgcgtccg gcgccgatca tttggtcatc
5101 ctgaccactg ccggaaaggt ctttaccgtg ggctgcgctg agcagggaca actcgggcgg
5161 ctgtcggagc gttccatctc cggagagggt cgtcgtggca aacgcgactt gctgcggccc
5221 acccagctca tcatcacgcg ggccaaacct tttgaggcca tctgggccac caactactgc
5281 acattcatgc gcgagtcaca gacacaggtc atctgggcca cagggcttaa caacttcaag
5341 cagctggccc acgagacaaa gggcaaggag ttcgcactga cgcccatcaa gacggaactc
5401 aaagatatcc gacatattgc cgggggtcag caccatactg tcatcctgac aacggacctt
5461 aagtgttccg tagtgggtcg tccggagtac ggtcggcttg gtctgggcga tgtcaaggat
5521 gttgtggaga agccaacgat tgtaaagaag ctgaccgaaa agattgtgtc cgttggatgt
5581 ggcgaagtct gttcctatgc cgttacaatt gatggcaaac tgtattcctg gggctccggt
5641 gtcaacaatc aactaggcgt tggcgacggc gatgatgaac tcgaaccgat cgtggttgtc
5701 agcaagaaca cgcaggggaa gcacatgctg ctcgcatctg gcggtggtca gcacgcaatc
5761 tttttggtca aagcggacaa acaggaccag gccggctaat gtacaacgcg tctcgagaga
5821 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
5881 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
5941 aactatatcc gactagtagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt
6001 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
6061 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
6121 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta
6181 aaagacgcag aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc
6241 ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa
6301 gttctgctat gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga
6361 cgaatacact attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt
6421 acggacggaa tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact
6481 gcggcgaact tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac
6541 aatatgggag accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata
6601 ccaaacgacg agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta
6661 ttaactggcg aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg
6721 gataaagttg caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat
6781 aaatctgggg caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt
6841 aagccttccc gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga
6901 aatagacaga ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt
6961 aaaggctcct tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt
7021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
7081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
7141 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
7201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
7261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
7321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
7381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
7441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
7501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
7561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
7621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
7681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
7741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
7801 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
7861 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
7921 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
7981 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
8041 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
8101 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
8161 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
8221 ttggtcactg atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga
8281 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
8341 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
8401 gtcaatgcca gcgcttcgtt aatacagatg taggtgtt
//
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