| 质粒名称: | pUC18-mini-Tn7T-Gm-lacZ |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 63120 |
| 存储实验室: | Herbert Schweizer |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18T-miniTn7T-Gm |
| 载体出品公司: | -- |
| 载体抗性: | 庆大霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 在miniTn7元素中插入无启动子lacZ |
| 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: | -- |
| 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: | pUC18-mini-Tn7T-Gm-lacZ was a gift from Herbert Schweizer (Addgene plasmid # 63120 ; http://n2t.net/addgene:63120 ; RRID:Addgene_63120) |
| 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 |
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 pUC18-mini-Tn7T-Gm-lacZ 7653 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION mini-Tn7 lacZ transcriptional fusion vector.
ACCESSION .
VERSION .
KEYWORDS pUC18-mini-Tn7T-Gm-lacZ
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7653)
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 7653)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7653
/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 958..1005
/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(1135..1668)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=GmR
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(1857..1885)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
protein_bind 1926..1973
/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)."
primer_bind 2231..2247
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2231..2247
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 2243..5317
/codon_start=1
/gene="lacZ"
/product="beta-galactosidase"
/label=lacZ
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
primer_bind complement(2263..2280)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2263..2279)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2272..2294)
/label=M13/pUC Forward
/note="In lacZ gene"
mobile_element 5399..5564
/mobile_element_type="transposon:Tn7"
/note="mini-Tn7 element (left end of the Tn7 transposon)"
rep_origin complement(5834..6422)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5914..5933)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6593..7453)
/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 7216..7235
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7454..7558)
/gene="bla"
/label=AmpR promoter
primer_bind 7626..7644
/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 atcgaattgg ggatcttgaa
961 gtacctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
1021 ctgatgtgct taaaaactta ctcaatggaa ttctagatcg aattggccgc ggcgttgtga
1081 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
1141 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
1201 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
1261 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
1321 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacttggg
1381 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
1441 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
1501 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
1561 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
1621 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
1681 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
1741 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
1801 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
1861 tacgaaccga acaggcttat gtcaattcga tccattgctg ttgacaaagg gaatcagggg
1921 atcttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttcagagcgc
1981 ttttgaagct aattcgatca tgcatgagct cactagtgga tcccccgggc tgcaggaatt
2041 cctcgagaag cttgggcccg gtacctcgcg aagggatcct aactaactag cgatcccgac
2101 tcactataga gggacaaact caaggtcatt cgcaagagtg gcctttatga ttgaccttct
2161 tccggttaat acgaccggga tcgagatcct aggtaggtag gggcgcggca ttttaacttt
2221 ctttatcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
2281 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
2341 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
2401 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
2461 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
2521 gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc
2581 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
2641 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttaactcgg cgtttcatct
2701 gtggtgcaac gggcgctggg tcggttacgg ccaggacagt cgtttgccgt ctgaatttga
2761 cctgagcgca tttttacgcg ccggagaaaa ccgcctcgcg gtgatggtgc tgcgttggag
2821 tgacggcagt tatctggaag atcaggatat gtggcggatg agcggcattt tccgtgacgt
2881 ctcgttgctg cataaaccga ctacacaaat cagcgatttc catgttgcca ctcgctttaa
2941 tgatgatttc agccgcgctg tactggaggc tgaagttcag atgtgcggcg agttgcgtga
3001 ctacctacgg gtaacagttt ctttatggca gggtgaaacg caggtcgcca gcggcaccgc
3061 gcctttcggc ggtgaaatta tcgatgagcg tggtggttat gccgatcgcg tcacactacg
3121 tctgaacgtc gaaaacccga aactgtggag cgccgaaatc ccgaatctct atcgtgcggt
3181 ggttgaactg cacaccgccg acggcacgct gattgaagca gaagcctgcg atgtcggttt
3241 ccgcgaggtg cggattgaaa atggtctgct gctgctgaac ggcaagccgt tgctgattcg
3301 aggcgttaac cgtcacgagc atcatcctct gcatggtcag gtcatggatg agcagacgat
3361 ggtgcaggat atcctgctga tgaagcagaa caactttaac gccgtgcgct gttcgcatta
3421 tccgaaccat ccgctgtggt acacgctgtg cgaccgctac ggcctgtatg tggtggatga
3481 agccaatatt gaaacccacg gcatggtgcc aatgaatcgt ctgaccgatg atccgcgctg
3541 gctaccggcg atgagcgaac gcgtaacgcg aatggtgcag cgcgatcgta atcacccgag
3601 tgtgatcatc tggtcgctgg ggaatgaatc aggccacggc gctaatcacg acgcgctgta
3661 tcgctggatc aaatctgtcg atccttcccg cccggtgcag tatgaaggcg gcggagccga
3721 caccacggcc accgatatta tttgcccgat gtacgcgcgc gtggatgaag accagccctt
3781 cccggctgtg ccgaaatggt ccatcaaaaa atggctttcg ctacctggag agacgcgccc
3841 gctgatcctt tgcgaatacg cccacgcgat gggtaacagt cttggcggtt tcgctaaata
3901 ctggcaggcg tttcgtcagt atccccgttt acagggcggc ttcgtctggg actgggtgga
3961 tcagtcgctg attaaatatg atgaaaacgg caacccgtgg tcggcttacg gcggtgattt
4021 tggcgatacg ccgaacgatc gccagttctg tatgaacggt ctggtctttg ccgaccgcac
4081 gccgcatcca gcgctgacgg aagcaaaaca ccagcagcag tttttccagt tccgtttatc
4141 cgggcaaacc atcgaagtga ccagcgaata cctgttccgt catagcgata acgagctcct
4201 gcactggatg gtggcgctgg atggtaagcc gctggcaagc ggtgaagtgc ctctggatgt
4261 cgctccacaa ggtaaacagt tgattgaact gcctgaacta ccgcagccgg agagcgccgg
4321 gcaactctgg ctcacagtac gcgtagtgca accgaacgcg accgcatggt cagaagccgg
4381 gcacatcagc gcctggcagc agtggcgtct ggcggaaaac ctcagtgtga cgctccccgc
4441 cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca tcgagctggg
4501 taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt ggattggcga
4561 taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc tggataacga
4621 cattggcgta agtgaagcga cccgcattga ccctaacgcc tgggtcgaac gctggaaggc
4681 ggcgggccat taccaggccg aagcagcgtt gttgcagtgc acggcagata cacttgctga
4741 tgcggtgctg attacgaccg ctcacgcgtg gcagcatcag gggaaaacct tatttatcag
4801 ccggaaaacc taccggattg atggtagtgg tcaaatggcg attaccgttg atgttgaagt
4861 ggcgagcgat acaccgcatc cggcgcggat tggcctgaac tgccagctgg cgcaggtagc
4921 agagcgggta aactggctcg gattagggcc gcaagaaaac tatcccgacc gccttactgc
4981 cgcctgtttt gaccgctggg atctgccatt gtcagacatg tataccccgt acgtcttccc
5041 gagcgaaaac ggtctgcgct gcgggacgcg cgaattgaat tatggcccac accagtggcg
5101 cggcgacttc cagttcaaca tcagccgcta cagtcaacag caactgatgg aaaccagcca
5161 tcgccatctg ctgcacgcgg aagaaggcac atggctgaat atcgacggtt tccatatggg
5221 gattggtggc gacgactcct ggagcccgtc agtatcggcg gagttccagc tgagcgccgg
5281 tcgctaccat taccagttgg tctggtgtca aaaataataa taaccgggca ggccatgtct
5341 gcccgtattt cgcgtaagga aatccattat gtactattta tcaagctcct tgcaggccaa
5401 ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta
5461 cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact
5521 ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt
5581 cctgttatgt ttgggcgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
5641 gtattgggcg ctcttccgct tcctcgctca ctgacccgct gcgctcggtc gttcggctgc
5701 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
5761 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
5821 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
5881 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
5941 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
6001 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
6061 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
6121 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
6181 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
6241 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
6301 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
6361 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
6421 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
6481 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
6541 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
6601 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
6661 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
6721 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
6781 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
6841 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
6901 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
6961 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
7021 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
7081 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
7141 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
7201 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
7261 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
7321 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
7381 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
7441 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
7501 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
7561 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
7621 ataaaaatag gcgtatcacg aggccctttc gtc
//
