| 质粒名称: | pTnsABC |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 160031 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCOLADuet-1 |
| 完整质粒大小: | 6858bp |
| 载体类型: | Bacterial Expression, CRISPR |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | VchTnsA, VchTnsB, VchTnsC |
| 物种来源: | Vibrio cholera |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | unknown (unknown if destroyed) |
| 3’克隆位点: | unknown (unknown if destroyed) |
| 5’测序引物 | unknown |
|
文献引用方法 |
|
| 材料和方法部分: | pTnsABC was a gift from Sheng Yang (Addgene plasmid # 160031 ; http://n2t.net/addgene:160031 ; RRID:Addgene_160031) |
| 参考文献部分: | Multicopy chromosomal integration using CRISPR-associated transposases. Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S. ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073. 10.1021/acssynbio.0c00073 PubMed 32551502 |
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 pTnsABC 6858 bp ds-DNA circular SYN 11-NOV-2021
DEFINITION Expresses V. cholerae TnsABC.
ACCESSION .
VERSION .
KEYWORDS pTnsABC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6858)
AUTHORS Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian
Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S
TITLE Multicopy chromosomal integration using CRISPR-associated
transposases.
JOURNAL ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073.
PUBMED 32551502
REFERENCE 2 (bases 1 to 6858)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 11, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6858
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3..27
/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 2149..2160
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(3587..3605)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 3601..3648
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(3878..4693)
/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 4601..4620
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(4694..4785)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4803..5438)
/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(4948..4967)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5636..6718)
/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 6680..6699
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(6719..6796)
/gene="lacI"
/label=lacI promoter
primer_bind 6842..3
/label=T7
/note="T7 promoter, forward primer"
promoter 6842..2
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atggcgacaa gtttacctac gccctcagca attacgactt cggcgttaga
121 gtatgcattc catactcccg ctcgcaatct aacgaaatct cgcggaaaaa acattcatcg
181 ttatgtcagt gtaaagatga gtaagaggat tacggtagaa tctactctag agtgtgatgc
241 ctgctatcac tttgattttg agccaagtat tgttcgcttt tgcgctcaac cgattcgatt
301 tttatattat ctcaatggtc agtctcactc ctatgttcct gactttctag ttcaatttga
361 taccaacgag tttgttctat atgaagtaaa gtcagcttat gctaagaaca aacctgattt
421 tgatgttgaa tgggaggcga aagtaaaagc agcaactgaa ctagggctag aattggagct
481 tgttgaagag agtgatatta gggatacggt tgtattaaat aatcttaagc gcatgcatcg
541 ttatgcttcg aaagatgagc tgaataacgt acataactct ctcttaaaaa taataaagta
601 caatggcgcc caatctgcaa gatgcttggg agaacagttg ggtttaaaag gccgaactgt
661 tttaccaatt ttgtgcgatt tgctgtcaag gtgtttactc gatacacgtt tggataagcc
721 tctatctctt gaatctcgat ttgagttggc cagttatggc taagaaaggg ttctcaagtt
781 tccatagaaa agcagtctct tcccaagata cgcttgaatc gatagagctt gtctctagcg
841 ctaattgtct agaaagtgtt acgtatcaag atatatcagc atttcccgaa acaattgcgg
901 tagagattaa tttccgatta agcattcttc gctttttagc gcgaaagtgc gaaaccattg
961 tggccaaatc aattgaacca catcgtgtag agctacagca aaactatagt agaaaaatac
1021 ccagtgcaat aacgatatat cgatggtggc ttgcttttcg aaaatcagac tacaacccca
1081 ttagcttagc acctaatatc aaggatagag gtaatagaga gacaaaagtg tcaacagttg
1141 ttgattctat tatggaacag gcagttgaaa gagttatatc tggacgaaaa gtcaatgtta
1201 gctctgcata taaacgtgtt cgacgaaaag ttcgtcaata caatctaact catggaacga
1261 aatacacgta tcctaagtac gaatctgtaa gaaagcgagt aaaaaagaaa accccatttg
1321 agttattagc cgcagggaaa ggggagagag tagctaagag agagtttcgc cgaatgggaa
1381 aaaagatcct cacgtctagc gtgctagaga gggttgaaat agatcacact gtcgttgacc
1441 tttttgcagt acacgaagag tatcgaatcc cattgggccg accttggctt actcaattgg
1501 ttgattgtta cagtaaagct gtaatcggtt tttatttagg tttcgagcct cctagctatg
1561 tgtcggtttc ccttgcactt aagaatgcaa tacaacgcaa agatgactta atctcctcgt
1621 atgaatcgat cgagaatgaa tggctatgtt atggcatccc agacctactc gtaactgata
1681 atggtaaaga gtttttgtcg aaagcatttg atcaagcatg tgaatcacta ttgatcaatg
1741 tgcatcaaaa taaagttgag acgcccgaca acaaacctca tgttgaacgt aactacggga
1801 ctattaatac ttctctgtta gacgatttac ctgggaaatc cttcagccag taccttcaaa
1861 gagaagggta cgactctgtg ggagaagcta cccttacact caatgagatt agagaaattt
1921 acttaatttg gttggtggat atttatcata aaaaacccaa tcagagaggc actaattgtc
1981 ctaatgttgc gtggaaaaag ggttgtcaag aatgggaacc agaggagttc tctggttcta
2041 aagacgaatt agactttaaa tttgctattg ttgattacaa acaacttact aaagtaggga
2101 taactgtcta caaagaactg agttatagca atgaccgttt agctgaatat agagggaaga
2161 aaggaaacca taaagttcag ttcaagtata accctgagtg tatggcagtt atttgggtgt
2221 tggatgagga tatgaatgag tactttacag ttaatgcgat tgactacgaa tatgcaagta
2281 gagtatcact ttggcaacat aaatataaca tgaaatatca agcagaacta aattcagcag
2341 aatatgatga ggacaaggaa attgatgcag aaataaaaat tgaagaaatc gcagatcgtt
2401 caattgttaa gactaacaaa atcagagctc ggaggcgtgg cgctaggcat caagagaata
2461 gcgcaagggc taagtcaatc agtaatgcga acccggcctc gatacaaaaa catgaagatg
2521 aaatcgttag tgcagataat gacgattggg atattgatta tgtctgagaa atcgtcaaat
2581 gagtgaaacg cgtgaggctc gtatatcaag agctaaaagg gcatttgtat ccacaccgag
2641 cgttaggaaa atcttgagtt acatggatag atgtagagat ctatcagacc tagagtctga
2701 gcctacatgc atgatggtct atggtgcttc gggtgtaggt aaaacgaccg tcatcaagaa
2761 atacttaaat caaaacagaa gagagtccga agccgggggc gatataatac cggttttgca
2821 tattgagttg ccagacaatg cgaagccagt agatgcagca agggaattgc tggttgaaat
2881 gggtgacccg ctagcacttt atgaaactga cttagctaga ttgacgaaaa gactgactga
2941 attaatccct gcggtcggcg tgaagctgat tattatcgat gagttccaac atttggtgga
3001 agaaaggtca aatcgggttc ttacccaagt aggtaattgg ctaaaaatga tacttaacaa
3061 aacgaaatgt ccaattgtta tatttggtat gccatactca aaagttgtac tgcaagcaaa
3121 ctcgcaactt cacgggcgat tttccattca ggttgaactg cgccccttta gctaccaggg
3181 aggtagaggt gtatttaaaa cttttttgga ataccttgat aaagccctac cttttgaaaa
3241 acaggctggc ttagccaacg aaagtttgca gaaaaaattg tatgcattct ctcagggaaa
3301 catgcgttcg ttgagaaacc ttatttatca agcatctatc gaagcaattg ataatcagca
3361 tgagacgata accgaagaag atttcgtttt tgcatcgaag ttgacatcgg gcgataaacc
3421 caactcatgg aaaaatcctt ttgaggaggg tgttgaggta acagaagata tgttacgacc
3481 gccaccaaaa gatattggtt gggaggacta tttgagacat tcaaccccga gagtgagtaa
3541 accaggtaga aataaaaact ttttcgaata acctaggctg ctgccaccgc tgagcaataa
3601 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
3661 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
3721 tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt
3781 ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
3841 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
3901 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
3961 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
4021 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
4081 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
4141 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
4201 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
4261 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
4321 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
4381 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
4441 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
4501 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
4561 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
4621 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
4681 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
4741 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc atgctagcgc
4801 agaaacgtcc tagaagatgc caggaggata cttagcagag agacaataag gccggagcga
4861 agccgttttt ccataggctc cgcccccctg acgaacatca cgaaatctga cgctcaaatc
4921 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct gatggctccc
4981 tcttgcgctc tcctgttccc gtcctgcggc gtccgtgttg tggtggaggc tttacccaaa
5041 tcaccacgtc ccgttccgtg tagacagttc gctccaagct gggctgtgtg caagaacccc
5101 ccgttcagcc cgactgctgc gccttatccg gtaactatca tcttgagtcc aacccggaaa
5161 gacacgacaa aacgccactg gcagcagcca ttggtaactg agaattagtg gatttagata
5221 tcgagagtct tgaagtggtg gcctaacaga ggctacactg aaaggacagt atttggtatc
5281 tgcgctccac taaagccagt taccaggtta agcagttccc caactgactt aaccttcgat
5341 caaaccgcct ccccaggcgg ttttttcgtt tacagagcag gagattacga cgatcgtaaa
5401 aggatctcaa gaagatcctt tacggattcc cgacaccatc actctagatt tcagtgcaat
5461 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
5521 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
5581 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
5641 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
5701 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
5761 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
5821 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
5881 agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg
5941 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
6001 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
6061 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
6121 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
6181 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
6241 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
6301 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
6361 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
6421 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
6481 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
6541 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
6601 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
6661 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
6721 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
6781 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa
6841 ttaatacgac tcactata
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pAraQCas-BsaI
- 下一篇：pHCas9-Nours
