| 质粒名称: | pUC18R6KT-mini-Tn7T-Km |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 64969 |
| 存储实验室: | Herbert Schweizer |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18R6KT-mini-Tn7T |
| 载体出品公司: | Herbert Schweizer |
| 载体抗性: | 氨苄青霉素和卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 添加FRT侧翼KmR标记 |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pUC18R6KT-mini-Tn7T-Km was a gift from Herbert Schweizer (Addgene plasmid # 64969 ; http://n2t.net/addgene:64969 ; RRID:Addgene_64969) |
| 参考文献部分: | A Tn7-based broad-range bacterial cloning and expression system. Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer RR, Schweizer HP. Nat Methods. 2005 Jun;2(6):443-8. 10.1038/nmeth765 PubMed 15908923 |
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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 pUC18R6KT-mini-Tn7T-Km 4911 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION mini-Tn7 base vector with transcriptional terminator and oriT.
ACCESSION .
VERSION .
KEYWORDS pUC18R6KT-mini-Tn7T-Km
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4911)
AUTHORS Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer
RR, Schweizer HP
TITLE A Tn7-based broad-range bacterial cloning and expression system.
JOURNAL Nat Methods. 2005 Jun;2(6):443-8.
PUBMED 15908923
REFERENCE 2 (bases 1 to 4911)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4911
/organism="synthetic DNA construct"
/mol_type="other DNA"
mobile_element 369..534
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
protein_bind complement(649..696)
/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)."
CDS 1083..1877
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(1137..1156)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1747..1766
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
protein_bind complement(1985..2032)
/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)."
terminator 2080..2166
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2269..2363
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(2391..2407)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2392..2408)
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind complement(2822..2841)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2941..2963
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3001..3019)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3087..3191
/gene="bla"
/label=AmpR promoter
CDS 3192..4052
/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(3410..3429)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4461..4817
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
oriT 4868..65
/note="incP origin of transfer"
ORIGIN
1 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
61 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
121 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
181 agggttatgc agcggaaagt ataccttaag gaatcccctg ataccgctcg ccgcagccga
241 acgaccgagc gcagcgggtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
301 cctctccccg cgcgttggcc gattcattaa tgcagcgccc aaacataaca ggaagaaaaa
361 tgccccgctg tgggcggaca aaatagttgg gaactgggag gggtggaaat ggagttttta
421 aggattattt agggaagagt gacaaaatag atgggaactg ggtgtagcgt cgtaagctaa
481 tacgaaaatt aaaaatgaca aaatagtttg gaactagatt tcacttatct ggttggcctg
541 caaggccttc gcgaggtacc gggcccaagc ttctcgagga attcctgcag cccgggggat
601 ccactagtga gctcatgcat gatctcgaat tagcttcaaa agcgctctga agttcctata
661 ctttctagag aataggaact tcggaatagg aacttcaaga tcccctgatt ccctttgtca
721 acagcaatgg ataattcgat ttaacaaatg catggcgcaa gggctgctaa aggaagcgga
781 acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
841 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
901 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
961 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
1021 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc
1081 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
1141 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
1201 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
1261 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
1321 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
1381 aggatctcct gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
1441 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
1501 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
1561 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
1621 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
1681 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
1741 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
1801 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
1861 ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagg atccaacatt tccaatcact
1921 agtgaattat ctagaattat tccattgagt aagtttttaa gcacatcagc ttcaaaagcg
1981 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggtact tcaagatccc
2041 caattcgaga tcgtccgggc cgcaagctcc tagcggcgga tttgtcctac tcaggagagc
2101 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
2161 tatttgatgc ctcaagctag agagtcatta ccccaggcgt ttaagggcac caataactgc
2221 cttaaaaaaa ttacgccccg ccctgccact catcgcagtc tagcttggat tctcaccaat
2281 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
2341 tactggatct atcaacagga gtccaagctc agctaattaa gctagcttat cgataccgtc
2401 gacctcgaac cccacgcccc tctttaatac gacgggcaat ttgcacttca gaaaatgaag
2461 agtttgcttt agccataaca aaagtccagt atgctttttc acagcataac tggactgatt
2521 tcagtttaca actattctgt ctagtttaag actttattgt catagtttag atctattttg
2581 ttcagtttaa gactttattg tccgcctaca cccgcttacg cagggcatcc atttattact
2641 caaccgtaac cgattttgcc aggttacgcg gctggtcctc tagctggcgt aatagcgaag
2701 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
2761 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
2821 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
2881 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
2941 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
3001 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
3061 caggtggcac ttttcgggga aatgtacgcg gaacccctat ttgtttattt ttctaaatac
3121 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
3181 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
3241 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
3301 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
3361 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
3421 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
3481 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
3541 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
3601 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
3661 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
3721 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
3781 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
3841 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
3901 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
3961 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
4021 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
4081 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
4141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
4201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
4261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
4321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
4381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
4441 taatcctgtt accagccggt tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt
4501 tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta
4561 aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt
4621 agaggctatt taagttgctg atttatatta attttattgt tcaaacatga gagcttagta
4681 cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt
4741 ttagttcgtt aaacatgaga gcttagtacg ttaaacttga gagcttagta cgtgaaacat
4801 gagagcttag tacgtactat caacaggttg aactgcccat gttctttcct gcgttatcag
4861 agcttatcgg ccagcctcgc agagcaggat tcccgttgag caccgccagg t
//
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