| 质粒名称: | pET-45b(+)-Tn5 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 112112 |
| 存储实验室: | Frank Pugh |
|
载体骨架基本信息 |
|
| 载体名称: | pET-45b(+) |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5260 bp |
| 完整质粒大小: | 6703 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: | Tn5 |
| 物种来源: | -- |
| 插入大小: | 1443 bp |
| 基因突变: | E54K E110K P242A L372P |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | His6（主干上的N终端） |
| 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: | pET-45b(+)-Tn5 was a gift from Frank Pugh (Addgene plasmid # 112112 ; http://n2t.net/addgene:112112 ; RRID:Addgene_112112) |
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
-Tn5质粒图谱.png)
载体简介
载体序列
LOCUS pET-45b(+)-Tn5 6646 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses hyperactive Tn5 transposase for next generation sequencing
library preparation.
ACCESSION .
VERSION .
KEYWORDS pET-45b(+)-Tn5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6646)
TITLE Bacterial expression plasmid for production of recombinant
hyperactive Tn5 transposase
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6646)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6646
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(99..118)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 309..330
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(582..1442)
/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 1205..1224
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1443..1535)
/gene="bla"
/label=AmpR promoter
rep_origin 1616..2204
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2105..2124
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2358..2375
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2390..2529
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2455..2474)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2615..2637
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2631..2822)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3394..4476)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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"
primer_bind 4438..4457
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4477..4554)
/gene="lacI"
/label=lacI promoter
primer_bind 4760..4779
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4867..4886
/label=T7
/note="T7 promoter, forward primer"
promoter 4867..4885
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 4886..4910
/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)."
RBS 4925..4947
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 4960..4977
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 6478..6522
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(6560..6578)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6574..6621
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
2401 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
2461 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
2521 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
2581 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
2641 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
2701 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
2761 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
2821 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
2881 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
2941 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
3001 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
3061 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
3121 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
3181 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
3241 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac
3301 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
3361 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
3421 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
3481 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
3541 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
3601 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
3661 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
3721 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
3781 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
3841 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
3901 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
3961 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
4021 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
4081 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
4141 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
4201 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
4261 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
4321 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
4381 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
4441 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
4501 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
4561 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
4621 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
4681 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
4741 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
4801 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
4861 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
4921 taattttgtt taactttaag aaggagatat accatggcac atcaccacca ccatcacgtg
4981 ggtaccatga ttacctcagc tctgcaccgc gcagccgatt gggcgaaaag cgtcttctcc
5041 tcagccgcac tgggcgatcc gcgtcgtacc gcacgtctgg ttaacgtcgc ggcccagctg
5101 gcgaaatata gtggtaaatc cattaccatc agctctgaag gtagcaaagc gatgcaagaa
5161 ggcgcctacc gttttattcg caacccgaat gtttctgcag aagctatccg caaagcgggc
5221 gccatgcaga ccgtcaaact ggcgcaagaa ttcccggaac tgctggccat tgaagatacc
5281 acgagtctgt cctatcgtca tcaggtggca gaaaaactgg gcaaactggg tagtatccaa
5341 gacaaatccc gcggttggtg ggtccatagc gtgctgctgc tggaagcgac cacgtttcgt
5401 accgttggcc tgctgcacca ggaatggtgg atgcgcccgg atgacccggc agatgctgat
5461 gaaaaagaaa gcggtaaatg gctggcagct gccgcaacgt cgcgtctgcg catgggctca
5521 atgatgtcga acgttattgc ggtctgcgat cgtgaagcgg acatccatgc ctatctgcag
5581 gataaactgg cccacaatga acgtttcgtg gttcgcagta aacatccgcg caaagatgtg
5641 gaatccggtc tgtatctgta cgaccacctg aaaaaccaac cggaactggg cggttaccag
5701 attagtatcg cccaaaaagg tgtcgtggat aaacgtggca aacgcaaaaa tcgtccggca
5761 cgcaaagctt cactgtcgct gcgtagcggt cgcattaccc tgaaacaggg caacatcacg
5821 ctgaatgcgg ttctggccga agaaattaac ccgccgaaag gcgaaacccc gctgaaatgg
5881 ctgctgctga cgtcagaacc ggttgaaagc ctggcccagg cactgcgtgt gattgatatc
5941 tacacccatc gttggcgcat tgaagaattt cacaaagcct ggaaaacggg cgcaggtgct
6001 gaacgtcagc gcatggaaga accggacaat ctggaacgta tggttagcat cctgtctttt
6061 gtggcagttc gtctgctgca actgcgcgaa tcctttaccc cgccgcaggc actgcgtgct
6121 caaggtctgc tgaaagaagc agaacacgtg gaaagccagt ctgctgaaac cgttctgacg
6181 ccggatgaat gtcaactgct gggctatctg gacaaaggta aacgtaaacg caaagaaaaa
6241 gcaggctctc tgcagtgggc ttacatggcg attgcccgtc tgggcggttt catggatagc
6301 aaacgcaccg gcatcgcatc ttggggtgct ctgtgggaag gctgggaagc gctgcagagc
6361 aaactggatg gcttcctggc ggcaaaagac ctgatggctc aaggcattaa aatctaagag
6421 ctcgtgtaca cggcgcgcct gcaggtcgac aagcttgcgg ccgcactcga gtctggtaaa
6481 gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag cgcagcttaa
6541 ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
6601 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggat
//
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