| 质粒名称: | pUC18T-mini-Tn7T-Zeo-gfpmut3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 65037 |
| 存储实验室: | Herbert Schweizer |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18T-mini-Tn7T-Zeo |
| 载体出品公司: | Herbert Schweizer |
| 载体抗性: | 氨苄青霉素和博来霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 5437 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | gfpmut3,GFP |
| 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: | -- |
| 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: | pUC18T-mini-Tn7T-Zeo-gfpmut3 was a gift from Herbert Schweizer (Addgene plasmid # 65037 ; http://n2t.net/addgene:65037 ; RRID:Addgene_65037) |
| 参考文献部分: | mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. Choi KH, Schweizer HP. Nat Protoc. 2006;1(1):153-61. 10.1038/nprot.2006.24 PubMed 17406227 |
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 pUC18T-mini-Tn7T-Zeo-gfpmut3 5430 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION ZeoR mini-Tn7 vector, mobilizable, for GFP tagging bacteria.
ACCESSION .
VERSION .
KEYWORDS pUC18T-mini-Tn7T-Zeo-gfpmut3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5430)
AUTHORS Choi KH, Schweizer HP
TITLE mini-Tn7 insertion in bacteria with single attTn7 sites: example
Pseudomonas aeruginosa.
JOURNAL Nat Protoc. 2006;1(1):153-61.
PUBMED 17406227
REFERENCE 2 (bases 1 to 5430)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5430
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 584..600
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 585..601
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 629..723
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator 826..912
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind complement(966..1013)
/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 complement(1096..1470)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(1489..1536)
/label=EM7 promoter
/note="synthetic bacterial promoter "
protein_bind complement(1647..1694)
/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 1748..1834
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1937..2031
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2051..2767)
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(2094..2114)
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
primer_bind 2709..2737
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
RBS 2774..2785
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
mobile_element 2947..3112
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
oriT 3271..3379
/note="incP origin of transfer"
rep_origin complement(3611..4199)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3691..3710)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4370..5230)
/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 4993..5012
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5231..5335)
/gene="bla"
/label=AmpR promoter
primer_bind 5403..5421
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
361 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc
601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattag cttcaaaagc
961 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac ttcaagatcc
1021 cctgattccc tttgtcaaca gcaatggatt tatctctgat gttacattgc acaagataaa
1081 aatatatcat catgatcagt cctgctcctc ggccacgaag tgcacgcagt tgccggccgg
1141 gtcgcgcagg gcgaactccc gcccccacgg ctgctcgccg atctcggtca tggccggccc
1201 ggaggcgtcc cggaagttcg tggacacgac ctccgaccac tcggcgtaca gctcgtccag
1261 gccgcgcacc cacacccagg ccagggtgtt gtccggcacc acctggtcct ggaccgcgct
1321 gatgaacagg gtcacgtcgt cccggaccac accggcgaag tcgtcctcca cgaagtcccg
1381 ggagaacccg agccggtcgg tccagaactc gaccgctccg gcgacgtcgc gcgcggtgag
1441 caccggaacg gcactggtca acttggccat ggtttagttc ctcaccttgt cgtattatac
1501 tatgccgata tactatgccg atgattaatt gtcaacacgt gctgatcatg accaaaatcc
1561 cttaacgtga gttacgcgtc gaagccgaat atctagaatt attccattga gtaagttttt
1621 aagcacatca gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc
1681 ggaataggta cttcaagatc cccaattcga tcatgcatga gctcgggccg caagctccta
1741 gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
1801 cccagtcttt cgactgagcc tttcgtttta tttgatgcct caagctagag agtcattacc
1861 ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc ctgccactca
1921 tcgcagtcta gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
1981 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagctcag
2041 ctaattaagc ttatttgtat agttcatcca tgccatgtgt aatcccagca gctgttacaa
2101 actcaagaag gaccatgtgg tctctctttt cgttgggatc tttcgaaagg gcagattgtg
2161 tggacaggta atggttgtct ggtaaaagga cagggccatc gccaattgga gtattttgtt
2221 gataatggtc tgctagttga acgcttccat cttcaatgtt gtgtctaatt ttgaagttaa
2281 ctttgattcc attcttttgt ttgtctgcca tgatgtatac attgtgtgag ttatagttgt
2341 attccaattt gtgtccaaga atgtttccat cttctttaaa atcaatacct tttaactcga
2401 ttctattaac aagggtatca ccttcaaact tgacttcagc acgtgtcttg tagttcccgt
2461 catctttgaa aaatatagtt ctttcctgta cataaccttc gggcatggca ctcttgaaaa
2521 agtcatgctg tttcatatga tctgggtatc tcgcaaagca ttgaacacca taaccgaaag
2581 tagtgacaag tgttggccat ggaacaggta gttttccagt agtgcaaata aatttaaggg
2641 taagttttcc gtatgttgca tcaccttcac cctctccact gacagaaaat ttgtgcccat
2701 taacatcacc atctaattca acaagaattg ggacaactcc agtgaaaagt tcttctcctt
2761 tacgcatgct taatttctcc tctttaattc tagagtggtg tcgcattata gggagttatt
2821 ccggcctgac tagaggaaat ttaaaataat tttctgaccg cgcaacgagc tcactagtgg
2881 atcccccggg ctgcaggaat tcctcgagaa gcttgggccc ggtacctcgc gaaggccttg
2941 caggccaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
3001 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
3061 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
3121 tttttcttcc tgttatgttt gggcgctgca ttaatgaatc ggccaacgcg cggggagagg
3181 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gacccgctgc gctcggtcgt
3241 tcggctgcgg cgagcggtat cagagcttat cggccagcct cgcagagcag gattcccgtt
3301 gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc
3361 tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac
3421 cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat
3481 aatgaccccg aagcagggtt atgcagcgga aagtatacct taaggaatcc cctgataacg
3541 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
3601 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcga
3661 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
3721 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
3781 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
3841 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
3901 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
3961 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
4021 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
4081 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
4141 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
4201 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
4261 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
4321 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
4381 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
4441 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
4501 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
4561 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
4621 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
4681 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
4741 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
4801 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
4861 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
4921 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
4981 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
5041 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
5101 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
5161 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
5221 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
5281 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
5341 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
5401 aaaataggcg tatcacgagg ccctttcgtc
//
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