| 质粒名称: | pMDISI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 51653 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pMD18-T Simple |
| 载体出品公司: | Takara |
| 空载体大小: | 2600bp |
| 完整质粒大小: | 4146 bp |
| 载体类型: | genetic modification |
| 载体抗性: | Ampicillin and Spectinomycin, 100 & 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | spcR |
| 插入大小: | 1500 bp |
| 启动子: | 未知 |
| 克隆方法: | TOPO Cloning |
| 5’测序引物 | M13F |
| 3’测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | pMDISI was a gift from Sheng Yang (Addgene plasmid # 51653 ; http://n2t.net/addgene:51653 ; RRID:Addgene_51653) |
| 参考文献部分: | High-Efficiency Scarless Genetic Modification in Escherichia coli by Using Lambda Red Recombination and I-SceI Cleavage. Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu J, Jiang W, Yang S. Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr 18. 10.1128/AEM.00313-14 PubMed 24747889 |
买家导航
pMDISI载体质粒基本信息
pMDISI质粒图谱载体图谱和pMDISI载体序列质粒序列多克隆位点信息
pMDISI质粒载体简介
pMDISI质粒序列载体序列
LOCUS pMDISI 4146 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION E coli genetic modification.
ACCESSION .
VERSION .
KEYWORDS pMDISI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4146)
AUTHORS Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu
J, Jiang W, Yang S
TITLE High-Efficiency Scarless Genetic Modification in Escherichia coli by
Using Lambda Red Recombination and I-SceI Cleavage.
JOURNAL Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr
18.
PUBMED 24747889
REFERENCE 2 (bases 1 to 4146)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4146
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..27)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(10..26)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(19..41)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(235..254)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 354..376
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(414..432)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 500..604
/gene="bla"
/label=AmpR promoter
CDS 605..1465
/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(823..842)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1636..2224
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2125..2144
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2378..2395
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2512..2533
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2548..2578
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2586..2602
/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 2591..2613
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2610..2626
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2610..2626
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
protein_bind 2721..2754
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
oriT 2761..2870
/note="incP origin of transfer"
protein_bind 4038..4084
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
1 ttctttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
61 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
121 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
181 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
241 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
301 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
361 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
421 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
841 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
1021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
1081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
1141 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
1201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
1261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
1321 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
1381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
1441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
1501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
1561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
1621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
1681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
1741 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
1801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
1861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
1921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
1981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
2041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
2101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
2161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
2221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
2281 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
2341 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
2401 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
2461 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
2521 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
2581 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgaact
2641 cgtcctcggt tcccggcgat cctctggaga ttatattacc ctgttatccc taattccggg
2701 gatccgtcga cctgcagttc gaagttccta ttctctagaa agtataggaa cttcgaagtt
2761 cccgccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag
2821 tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc
2881 gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga
2941 aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg
3001 aaaatgcagc tcacggtaac tgatgccgta tttgcagtac cagcgtacgg cccacagaat
3061 gatgtcacgc tgaaaatgcc ggcctttgaa tgggttcatg tgcagctcca taagcaaaag
3121 gggatgataa gtttatcacc accgactatt tgcaacagtg ccgttgatcg tgctatgatc
3181 gactgatgtc atcagcggtg gagtgcaatg tcatgaggga agcggtgatc gccgaagtat
3241 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
3301 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
3361 tgctggttac ggtgaccgta aggcttgatg aaaacaacgc ggcgagcttt gatcaacgac
3421 ctttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
3481 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
3541 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
3601 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
3661 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
3721 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
3781 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
3841 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
3901 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
3961 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataagatgcc gctcgccagt
4021 cgattggctg agctcataag ttcctattcc gaagttccta ttctctagaa agtataggaa
4081 cttcgaagca gctccagcct acacattacc ctgttatccc taataatcgt ccacttgccg
4141 tcatgc
//
