| 质粒名称: | pPSU150 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 173836 |
| 存储实验室: | Song Tan |
|
载体骨架基本信息 |
|
| 载体名称: | pST50Tr |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 6808 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | STRHSTPABMBPpepN |
| 物种来源: | Synthetic |
| 启动子: | T7 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NdeI (unknown if destroyed) |
| 3’克隆位点: | BsrGI (unknown if destroyed) |
| 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: | pPSU150 was a gift from Song Tan (Addgene plasmid # 173836 ; http://n2t.net/addgene:173836 ; RRID:Addgene_173836) |
| 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 pPSU150 6808 bp ds-DNA circular SYN 17-AUG-2021
DEFINITION Expresses 150 kD Penn State ladder protein.
ACCESSION .
VERSION .
KEYWORDS pPSU150
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6808)
AUTHORS Tan, S
TITLE TanLab Protein Ladder Plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 6808)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 17, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6808
/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 113..135
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 152..175
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 185..214
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 413..1510
/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 1367..1376
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1484..1507
/label=MBP-F
/note="Maltose binding protein, forward primer"
primer_bind complement(4240..4258)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4254..4301
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 4366..4457
/gene="bla"
/label=AmpR promoter
terminator 5322..5361
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
rep_origin 5392..5980
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5881..5900
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6134..6151
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 6166..6305
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(6231..6250)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6391..6413
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(6407..6598)
/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 tccctccggc tccggagagc tccaattgga attcgctagc tctagaaata attttgttta
121 actttaagaa ggagatatac atatggcaag ctggtcacat ccgcagtttg aaaaaggtag
181 cagtcatcat caccaccatc accatcatca tcacagcagc ggtagcggta gcgcagataa
241 caaatttaac aaagaacagc agaacgcctt ctacgaaatt ctgcatctgc cgaatctgaa
301 tgaagaacag cgtaatggtt ttatccagag cctgaaagat gatccgagcc agagcgcaaa
361 tctgctggcc gaagcaaaaa aactgaatga tgcacaggca ccgaaaggat ctaaaatcga
421 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt
481 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact
541 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc
601 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa
661 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct
721 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc
781 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg
841 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc
901 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt
961 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca
1021 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc
1081 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg
1141 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag
1201 cgcaggtatt aacgccgcca gtccgaacaa agagctggcg aaagagttcc tcgaaaacta
1261 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc
1321 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa
1381 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt
1441 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga
1501 cgcgcagact aactctggat ccactcaaca gccacaagcc aaataccgtc acgattatcg
1561 tgcgccggat taccagatta ctgatattga cttgaccttt gacctcgacg cgcaaaagac
1621 ggtcgttacc gcggtcagcc aggctgtccg tcatggtgca tcagatgctc cccttcgtct
1681 caacggcgaa gacctcaaac tggtttctgt tcatattaat gatgagccgt ggaccgcctg
1741 gaaagaagaa gagggcgcac tggttatcag taatttgccg gagcgtttta cgcttaagat
1801 cattaatgaa ataagcccgg cggcgaatac cgcgctggaa gggctttatc agtcaggcga
1861 tgcgctttgc acccagtgtg aagccgaagg tttccgccat attacgtatt atctcgaccg
1921 cccggacgtg ctggcgcgtt ttaccaccaa aattattgcc gataaaatca aatatccctt
1981 cctgctttcc aatggtaacc gcgttgcgca aggcgaactg gaaaacggac gccattgggt
2041 acagtggcag gacccgttcc cgaaaccgtg ctacctgttt gcgctggtgg caggcgactt
2101 tgatgtactg cgcgatacct ttaccacgcg ttctggtcgc gaagtagcac tggagctgta
2161 cgtcgatcgc ggcaaccttg atcgcgcgcc gtgggcgatg acctcgctga aaaactccat
2221 gaaatgggat gaagaacgct ttggcctgga gtatgacctc gacatctata tgatcgtcgc
2281 ggtggatttc ttcaatatgg gcgcaatgga gaataagggg ctgaatatct ttaactccaa
2341 atatgtgctg gcccgcaccg acaccgccac cgacaaagat tacctcgata ttgaacgcgt
2401 tatcggccat gaatatttcc ataactggac cggtaaccga gtgacctgtc gcgactggtt
2461 ccagctcagc ctgaaagaag gtttaaccgt cttccgcgat caggagttca gctctgacct
2521 tggttcccgc gcagttaacc gcatcaataa tgtacgcacc atgcgcggat tgcagtttgc
2581 agaagacgcc agcccgatgg cgcacccgat ccgcccggat atggtcattg agatgaacaa
2641 cttctacacc ctgaccgttt acgagaaggg cgcggaagtg attcgcatga tccacaccct
2701 gcttggcgaa gaaaacttcc agaaagggat gcagctttat ttcgagcgtc atgatggtag
2761 tgcagcgacc tgtgacgact ttgtgcaggc gatggaagat gcgtcgaatg tcgatctctc
2821 ccatttccgc cgttggtaca gccagtccgg tacaccgatt gtgaccgtca aagacgacta
2881 caatccggaa accgagcagt acaccctgac catcagccag cgcacgccag ccacgccgga
2941 tcaggcagaa aaacagccgc tgcatattcc gtttgccatc gaactgtatg ataacgaagg
3001 caaagtgatc ccgttgcaga aaggcggtca tccggtgaat tccgtgctga acgtcactca
3061 ggcggaacag acctttgtct ttgataatgt ctacttccag ccggtgcctg cgctgctgtg
3121 cgaattctct gcgccagtga aactggaata taagtggagc gatcagcaac tgaccttcct
3181 gatgcgtcat gcgcgtaatg atttctcccg ctgggatgcg gcgcaaagtt tgctggcaac
3241 ctacatcaag ctgaacgtcg cgcgtcatca gcaaggtcag ccgctgtctc tgccggtgca
3301 tgtggctgat gctttccgcg cggtactgct tgatgagaag attgatccag cgctggcggc
3361 agaaatcctg acgctgcctt ctgtcaatga aatggctgaa ttgttcgata tcatcgaccc
3421 gattgctatt gccgaagtac gcgaagcact cactcgtact ctggcgactg aactggcgga
3481 tgagctactg gctatttaca acgcgaatta ccagagcgag taccgtgttg agcatgaaga
3541 tattgcaaaa cgcactctgc gtaatgcctg cctgcgtttc ctcgcttttg gtgaaacgca
3601 tctggctgat gtgctggtga gcaagcagtt ccacgaagca aacaatatga ctgatgcgct
3661 ggcggcgctt tctgcggcgg ttgccgcaca gctgccttgc cgtgacgcgc tgatgcagga
3721 gtacgacgac aagtggcatc agaacggtct ggtgatggat aaatggttta tcctgcaagc
3781 caccagcccg gcggcgaatg tgctggagac ggtgcgcggc ctgttgcagc atcgctcatt
3841 taccatgagc aacccgaacc gtattcgttc gttgattggc gcgtttgcgg gcagcaatcc
3901 ggcagcgttc catgccgaag atggcagcgg ttacctgttc ctggtggaaa tgcttaccga
3961 cctcaacagc cgtaacccgc aggtggcttc acgtctgatt gaaccgctga ttcgcctgaa
4021 acgttacgat gccaaacgtc aggagaaaat gcgcgcggcg ctggaacagt tgaaagggct
4081 ggaaaatctc tctggcgatc tgtacgagaa gataactaaa gcactggctc agtacactaa
4141 aaactaatgt acaagatcca gatctaagct tggtaccacg cgtgcgcgct gatccggctg
4201 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
4261 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
4321 cgggattagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt ttatttttct
4381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
4441 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
4501 cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta aaagacgcag
4561 aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc ggtaaaatcc
4621 ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa gttctgctat
4681 gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga cgaatacact
4741 attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt acggacggaa
4801 tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact gcggcgaact
4861 tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac aatatgggag
4921 accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata ccaaacgacg
4981 agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta ttaactggcg
5041 aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg gataaagttg
5101 caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat aaatctgggg
5161 caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt aagccttccc
5221 gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga aatagacaga
5281 ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt aaaggctcct
5341 tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt cttgagatcc
5401 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
5461 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
5521 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
5581 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
5641 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
5701 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
5761 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
5821 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
5881 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
5941 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
6001 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
6061 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
6121 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
6181 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
6241 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
6301 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
6361 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
6421 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
6481 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
6541 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
6601 atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga aacgagagag
6661 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
6721 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
6781 gcgcttcgtt aatacagatg taggtgtt
//
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