| 质粒名称: | pSL0283 (pTnsABC) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 130633 |
| 存储实验室: | Samuel H. Sternberg |
|
载体骨架基本信息 |
|
| 载体名称: | pCOLADuet-1 |
| 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，CRISPR；转座子 |
| 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: | VchTnsA, VchTnsB, VchTnsC |
| 物种来源: | Vibrio cholerae |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | WP_000202732.1, WP_001074608.1, WP_002024152.1 |
| 启动子 | T7 |
| 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: | pSL0283 (pTnsABC) was a gift from Samuel H. Sternberg (Addgene plasmid # 130633 ; http://n2t.net/addgene:130633 ; RRID:Addgene_130633) |
| 参考文献部分: | Transposon-encoded CRISPR-Cas systems direct RNA-guided DNA integration. Klompe SE, Vo PLH, Halpin-Healy TS, Sternberg SH. Nature. 2019 Jul;571(7764):219-225. doi: 10.1038/s41586-019-1323-z. Epub 2019 Jun 12. 10.1038/s41586-019-1323-z PubMed 31189177 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pSL0283 (pTnsABC) 6858 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses V. cholerae TnsA, TnsB, and TnsC from a T7 promoter..
ACCESSION .
VERSION .
KEYWORDS pSL0283 (pTnsABC)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6858)
AUTHORS Klompe SE, Vo PLH, Halpin-Healy TS, Sternberg SH
TITLE Transposon-encoded CRISPR-Cas systems direct RNA-guided DNA
integration.
JOURNAL Nature. 2019 Jul;571(7764):219-225. doi: 10.1038/s41586-019-1323-z.
Epub 2019 Jun 12.
PUBMED 31189177
REFERENCE 2 (bases 1 to 6858)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6858
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(198..1280)
/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 1242..1261
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(1281..1358)
/gene="lacI"
/label=lacI promoter
primer_bind 1404..1423
/label=T7
/note="T7 promoter, forward primer"
promoter 1404..1422
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1423..1447
/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)."
CDS 3569..3580
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5007..5025)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5021..5068
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5298..6113)
/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 6021..6040
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(6114..6205)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6223..6858)
/direction=LEFT
/label=ColA ori
/note="Plasmids containing the ColA origin of replication
can be propagated in E. coli cells that contain additional
plasmids with compatible origins."
primer_bind complement(6368..6387)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tcactctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
61 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
121 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
301 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
361 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
421 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
481 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
541 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
601 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
661 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
721 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
781 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
841 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
901 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
961 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
1021 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
1081 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
1141 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
1201 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
1261 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
1321 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
1381 atgcgactcc tgcattagga aattaatacg actcactata ggggaattgt gagcggataa
1441 caattcccct gtagaaataa ttttgtttaa ctttaataag gagatatacc atggcgacaa
1501 gtttacctac gccctcagca attacgactt cggcgttaga gtatgcattc catactcccg
1561 ctcgcaatct aacgaaatct cgcggaaaaa acattcatcg ttatgtcagt gtaaagatga
1621 gtaagaggat tacggtagaa tctactctag agtgtgatgc ctgctatcac tttgattttg
1681 agccaagtat tgttcgcttt tgcgctcaac cgattcgatt tttatattat ctcaatggtc
1741 agtctcactc ctatgttcct gactttctag ttcaatttga taccaacgag tttgttctat
1801 atgaagtaaa gtcagcttat gctaagaaca aacctgattt tgatgttgaa tgggaggcga
1861 aagtaaaagc agcaactgaa ctagggctag aattggagct tgttgaagag agtgatatta
1921 gggatacggt tgtattaaat aatcttaagc gcatgcatcg ttatgcttcg aaagatgagc
1981 tgaataacgt acataactct ctcttaaaaa taataaagta caatggcgcc caatctgcaa
2041 gatgcttggg agaacagttg ggtttaaaag gccgaactgt tttaccaatt ttgtgcgatt
2101 tgctgtcaag gtgtttactc gatacacgtt tggataagcc tctatctctt gaatctcgat
2161 ttgagttggc cagttatggc taagaaaggg ttctcaagtt tccatagaaa agcagtctct
2221 tcccaagata cgcttgaatc gatagagctt gtctctagcg ctaattgtct agaaagtgtt
2281 acgtatcaag atatatcagc atttcccgaa acaattgcgg tagagattaa tttccgatta
2341 agcattcttc gctttttagc gcgaaagtgc gaaaccattg tggccaaatc aattgaacca
2401 catcgtgtag agctacagca aaactatagt agaaaaatac ccagtgcaat aacgatatat
2461 cgatggtggc ttgcttttcg aaaatcagac tacaacccca ttagcttagc acctaatatc
2521 aaggatagag gtaatagaga gacaaaagtg tcaacagttg ttgattctat tatggaacag
2581 gcagttgaaa gagttatatc tggacgaaaa gtcaatgtta gctctgcata taaacgtgtt
2641 cgacgaaaag ttcgtcaata caatctaact catggaacga aatacacgta tcctaagtac
2701 gaatctgtaa gaaagcgagt aaaaaagaaa accccatttg agttattagc cgcagggaaa
2761 ggggagagag tagctaagag agagtttcgc cgaatgggaa aaaagatcct cacgtctagc
2821 gtgctagaga gggttgaaat agatcacact gtcgttgacc tttttgcagt acacgaagag
2881 tatcgaatcc cattgggccg accttggctt actcaattgg ttgattgtta cagtaaagct
2941 gtaatcggtt tttatttagg tttcgagcct cctagctatg tgtcggtttc ccttgcactt
3001 aagaatgcaa tacaacgcaa agatgactta atctcctcgt atgaatcgat cgagaatgaa
3061 tggctatgtt atggcatccc agacctactc gtaactgata atggtaaaga gtttttgtcg
3121 aaagcatttg atcaagcatg tgaatcacta ttgatcaatg tgcatcaaaa taaagttgag
3181 acgcccgaca acaaacctca tgttgaacgt aactacggga ctattaatac ttctctgtta
3241 gacgatttac ctgggaaatc cttcagccag taccttcaaa gagaagggta cgactctgtg
3301 ggagaagcta cccttacact caatgagatt agagaaattt acttaatttg gttggtggat
3361 atttatcata aaaaacccaa tcagagaggc actaattgtc ctaatgttgc gtggaaaaag
3421 ggttgtcaag aatgggaacc agaggagttc tctggttcta aagacgaatt agactttaaa
3481 tttgctattg ttgattacaa acaacttact aaagtaggga taactgtcta caaagaactg
3541 agttatagca atgaccgttt agctgaatat agagggaaga aaggaaacca taaagttcag
3601 ttcaagtata accctgagtg tatggcagtt atttgggtgt tggatgagga tatgaatgag
3661 tactttacag ttaatgcgat tgactacgaa tatgcaagta gagtatcact ttggcaacat
3721 aaatataaca tgaaatatca agcagaacta aattcagcag aatatgatga ggacaaggaa
3781 attgatgcag aaataaaaat tgaagaaatc gcagatcgtt caattgttaa gactaacaaa
3841 atcagagctc ggaggcgtgg cgctaggcat caagagaata gcgcaagggc taagtcaatc
3901 agtaatgcga acccggcctc gatacaaaaa catgaagatg aaatcgttag tgcagataat
3961 gacgattggg atattgatta tgtctgagaa atcgtcaaat gagtgaaacg cgtgaggctc
4021 gtatatcaag agctaaaagg gcatttgtat ccacaccgag cgttaggaaa atcttgagtt
4081 acatggatag atgtagagat ctatcagacc tagagtctga gcctacatgc atgatggtct
4141 atggtgcttc gggtgtaggt aaaacgaccg tcatcaagaa atacttaaat caaaacagaa
4201 gagagtccga agccgggggc gatataatac cggttttgca tattgagttg ccagacaatg
4261 cgaagccagt agatgcagca agggaattgc tggttgaaat gggtgacccg ctagcacttt
4321 atgaaactga cttagctaga ttgacgaaaa gactgactga attaatccct gcggtcggcg
4381 tgaagctgat tattatcgat gagttccaac atttggtgga agaaaggtca aatcgggttc
4441 ttacccaagt aggtaattgg ctaaaaatga tacttaacaa aacgaaatgt ccaattgtta
4501 tatttggtat gccatactca aaagttgtac tgcaagcaaa ctcgcaactt cacgggcgat
4561 tttccattca ggttgaactg cgccccttta gctaccaggg aggtagaggt gtatttaaaa
4621 cttttttgga ataccttgat aaagccctac cttttgaaaa acaggctggc ttagccaacg
4681 aaagtttgca gaaaaaattg tatgcattct ctcagggaaa catgcgttcg ttgagaaacc
4741 ttatttatca agcatctatc gaagcaattg ataatcagca tgagacgata accgaagaag
4801 atttcgtttt tgcatcgaag ttgacatcgg gcgataaacc caactcatgg aaaaatcctt
4861 ttgaggaggg tgttgaggta acagaagata tgttacgacc gccaccaaaa gatattggtt
4921 gggaggacta tttgagacat tcaaccccga gagtgagtaa accaggtaga aataaaaact
4981 ttttcgaata acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
5041 ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt
5101 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
5161 acgaccctgc cctgaaccga cgacaagctg acgaccgggt ctccgcaagt ggcacttttc
5221 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
5281 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
5341 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
5401 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
5461 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
5521 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
5581 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
5641 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa
5701 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
5761 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
5821 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
5881 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
5941 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
6001 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
6061 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc
6121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
6181 gaatgtattt agaaaaataa acaaataggc atgctagcgc agaaacgtcc tagaagatgc
6241 caggaggata cttagcagag agacaataag gccggagcga agccgttttt ccataggctc
6301 cgcccccctg acgaacatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
6361 ggactataaa gataccaggc gtttccccct gatggctccc tcttgcgctc tcctgttccc
6421 gtcctgcggc gtccgtgttg tggtggaggc tttacccaaa tcaccacgtc ccgttccgtg
6481 tagacagttc gctccaagct gggctgtgtg caagaacccc ccgttcagcc cgactgctgc
6541 gccttatccg gtaactatca tcttgagtcc aacccggaaa gacacgacaa aacgccactg
6601 gcagcagcca ttggtaactg agaattagtg gatttagata tcgagagtct tgaagtggtg
6661 gcctaacaga ggctacactg aaaggacagt atttggtatc tgcgctccac taaagccagt
6721 taccaggtta agcagttccc caactgactt aaccttcgat caaaccgcct ccccaggcgg
6781 ttttttcgtt tacagagcag gagattacga cgatcgtaaa aggatctcaa gaagatcctt
6841 tacggattcc cgacacca
//
