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- PVT10577
- 2025年07月15日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
负20摄氏度
- 保质期：
3个月
- 英文名：
pET-24b
- 库存：
50
- 供应商：
LSMBIO
- 规格：
2ug
pET-24b
Catalog No. PVT10577
Packing 2ug
Function E.coli expression plasmid
Resistance Kan
Screen /
Strain DH5alpha
Culture temperature 37degrees centigrade
Replicon pBR322
Copy Low
Promoter T7
Induction IPTG
Forward primer T7
Reverse primer T7-ter
pET-24b Description
The pET-24a-d(+) vectors carry an N-terminal T7?Tag sequence plus an optional C-terminal His?Tag sequence. These vectors differ from pET-21a-d(+) only by their selectable marker (kanamycin vs. ampicillin resistance). Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, singlestranded sequencing should be performed using the T7 terminator primer.
pET-24b cloning site
pET-24b Sequence
LOCUS Exported 5309 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pET-24b
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5309)
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016
FEATURES Location/Qualifiers
source 1..5309
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(206..238)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
protein_bind 283..307
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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)."
promoter complement(308..326)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 635..712
/gene="lacI"
/note="lacI promoter"
CDS 713..1795
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS 2604..2795
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3225..3813)
/direction=LEFT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS 3935..4750
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(4843..5298)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tcccgaccca tttgctgtcc accagtcatg ctagccatat
241 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
301 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
361 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
421 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
481 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
541 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
601 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
661 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
721 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
781 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
841 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
901 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
961 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
1021 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
1081 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
1141 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
1201 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
1261 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
1321 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
1381 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
1441 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
1501 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
1561 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
1621 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
1681 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
1741 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
1801 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
1861 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
1921 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
1981 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
2041 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
2101 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
2161 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
2221 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
2281 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
2341 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
2401 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
2461 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
2521 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
2581 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
2641 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
2701 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
2761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
2821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
2881 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
2941 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
3001 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
3061 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
3121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
3181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
3241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
3301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
3361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
3421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
3481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
3541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
3601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
3661 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
3721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
3781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
3901 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
3961 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
4021 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
4081 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
4141 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
4201 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
4261 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
4321 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
4381 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
4441 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
4501 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
4561 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
4621 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
4681 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
4741 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
4801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
4861 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
4921 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
4981 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
5041 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
5101 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
5161 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
5221 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
5281 cgccgctaca gggcgcgtcc cattcgcca
//
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文献和实验This protocol is for Mini (up to 20 µg) preparations of high-copy plasmid DNA from cultures of E. coli . Important notes before starting New users are strongly recommended to read Appendix A (page 21) at the end
Plasmid Profile Typing and Plasmid Fingerprinting
Plasmids are extra-chromosomal molecules of deoxyribonucleic acid (DNA) capable of autonomous replication. Such molecules have been identified in many bacterial genera and usually exist as covalently closed circular (CCC) molecules. Plasmids
质粒 plasmid 在细胞内整个世代稳定地传递给后代，但并不在染色体上而进行自主增殖的一类遗传因子的总称。该类因子的存在与否，对细胞的生存无关紧要。这个名词主要指细菌细胞内的因子，它们包括可在细胞间发生传递的F因子、 R因子、大肠杆菌素因子等。此外，还有各种本身不能传递的因子。在具有质粒细胞的后代中，有时会出现低频率的失去质粒的细菌。在质粒中，对那些能变成插入奇主染色体状态的特殊质粒，特称为附加体（episome）。
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