| 质粒名称: | psfTn5 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79107 |
| 存储实验室: | Sten Linnarsson |
|
载体骨架基本信息 |
|
| 载体名称: | pNIC-Bsa4 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 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: | transposase 5,Tn5 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | E54K and L372P |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | His（主干上的N终端） |
| 克隆方法: | Ligation Independent克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T7 terminal |
|
文献引用方法 |
|
| 材料和方法部分: | psfTn5 was a gift from Sten Linnarsson (Addgene plasmid # 79107 ; http://n2t.net/addgene:79107 ; RRID:Addgene_79107) |
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 psfTn5 6787 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION bacterial expression of hyperactive Tn5 transposase carrying E54K
and L372P mutations.
ACCESSION .
VERSION .
KEYWORDS psfTn5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6787)
TITLE Linnarsson lab plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6787)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6787
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 92..109
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 134..154
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 155..1582
/codon_start=1
/product="transposase from the bacterial Tn5 transposon
(Reznikoff, 1993)"
/label=Tn5 transposase
/translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
KLDGFLAAKDLMAQGIKI"
CDS 1648..1665
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1718..1736)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1732..1779
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1816..2271
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1903..1922)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2113..2134
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(2364..3179)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 3087..3106
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3171..3190
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 3301..3889
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3790..3809
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4043..4060
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4075..4217
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4143..4162)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4303..4325
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4319..4510)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(5319..6401)
/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 6363..6382
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(6402..6479)
/gene="lacI"
/label=lacI promoter
primer_bind 6685..6704
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
121 agatctgggt accgagaacc tgtacttcca atccatgatt accagtgcac tgcatcgtgc
181 ggcggattgg gcgaaaagcg tgttttctag tgctgcgctg ggtgatccgc gtcgtaccgc
241 gcgtctggtg aatgttgcgg cgcaactggc caaatatagc ggcaaaagca ttaccattag
301 cagcgaaggc agcaaagcca tgcaggaagg cgcgtatcgt tttattcgta atccgaacgt
361 gagcgcggaa gcgattcgta aagcgggtgc catgcagacc gtgaaactgg cccaggaatt
421 tccggaactg ctggcaattg aagataccac ctctctgagc tatcgtcatc aggtggcgga
481 agaactgggc aaactgggta gcattcagga taaaagccgt ggttggtggg tgcatagcgt
541 gctgctgctg gaagcgacca cctttcgtac cgtgggcctg ctgcatcaag aatggtggat
601 gcgtccggat gatccggcgg atgcggatga aaaagaaagc ggcaaatggc tggccgctgc
661 tgcaacttcg cgtctgagaa tgggcagcat gatgagcaac gtgattgcgg tgtgcgatcg
721 tgaagcggat attcatgcgt atctgcaaga taaactggcc cataacgaac gttttgtggt
781 gcgtagcaaa catccgcgta aagatgtgga aagcggcctg tatctgtatg atcacctgaa
841 aaaccagccg gaactgggcg gctatcagat tagcattccg cagaaaggcg tggtggataa
901 acgtggcaaa cgtaaaaacc gtccggcgcg taaagcgagc ctgagcctgc gtagcggccg
961 tattaccctg aaacagggca acattaccct gaacgcggtg ctggccgaag aaattaatcc
1021 gccgaaaggc gaaaccccgc tgaaatggct gctgctgacc agcgagccgg tggaaagtct
1081 ggcccaagcg ctgcgtgtga ttgatattta tacccatcgt tggcgcattg aagaatttca
1141 caaagcgtgg aaaacgggtg cgggtgcgga acgtcagcgt atggaagaac cggataacct
1201 ggaacgtatg gtgagcattc tgagctttgt ggcggtgcgt ctgctgcaac tgcgtgaatc
1261 ttttactccg ccgcaagcac tgcgtgcgca gggcctgctg aaagaagcgg aacacgttga
1321 aagccagagc gcggaaaccg tgctgacccc ggatgaatgc caactgctgg gctatctgga
1381 taaaggcaaa cgcaaacgca aagaaaaagc gggcagcctg caatgggcgt atatggcgat
1441 tgcgcgtctg ggcggcttta tggatagcaa acgtaccggc attgcgagct ggggtgcgct
1501 gtgggaaggt tgggaagcgc tgcaaagcaa actggatggc tttctggccg cgaaagacct
1561 gatggcgcag ggcattaaaa tctgacagta aaggtggata cggatccgaa ttcgagctcc
1621 gtcgacaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct
1681 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
1741 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
1801 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
1861 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
1921 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
1981 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
2041 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
2101 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
2161 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
2221 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tcaggtggca
2281 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
2341 tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
2401 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
2461 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
2521 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
2581 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
2641 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
2701 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
2761 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
2821 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
2881 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
2941 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
3001 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
3061 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
3121 tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata
3181 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc
3241 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
3301 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
3361 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
3421 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
3481 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
3541 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
3601 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
3661 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
3721 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
3781 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
3841 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
3901 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
3961 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
4021 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
4081 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc
4141 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
4201 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
4261 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
4321 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
4381 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
4441 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
4501 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
4561 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
4621 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
4681 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
4741 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
4801 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
4861 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
4921 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
4981 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
5041 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
5101 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
5161 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
5221 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
5281 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
5341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
5401 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
5461 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
5521 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
5581 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
5641 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
5701 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
5761 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
5821 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
5881 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
5941 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
6001 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
6061 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
6121 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
6181 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
6241 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
6301 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
6361 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
6421 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
6481 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
6541 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
6601 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
6661 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
6721 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
6781 gcgaaat
//
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