| 质粒名称: | pTet-tns |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170628 |
| 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: | 6403 bp |
| 载体类型: | 细菌表达 |
| 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: | VchTnsABC |
| 物种来源: | Vibrio cholera |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pTet-tns was a gift from Sheng Yang (Addgene plasmid # 170628 ; http://n2t.net/addgene:170628 ; RRID:Addgene_170628) |
| 参考文献部分: | Programming Cells by Multicopy Chromosomal Integration Using CRISPR-Associated Transposases. Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J, Yang S. CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018. 10.1089/crispr.2021.0018 PubMed 34152213 |
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 pTet-tns 6403 bp ds-DNA circular SYN 03-AUG-2021
DEFINITION Expression of CRISPR-associated transposases.
ACCESSION .
VERSION .
KEYWORDS pTet-tns
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6403)
AUTHORS Zhang Y, Yang J, Yang S, Zhang J, Chen J, Tao R, Jiang Y, Yang J,
Yang S
TITLE Programming Cells by Multicopy Chromosomal Integration Using
CRISPR-Associated Transposases.
JOURNAL CRISPR J. 2021 Jun;4(3):350-359. doi: 10.1089/crispr.2021.0018.
PUBMED 34152213
REFERENCE 2 (bases 1 to 6403)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 3, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6403
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 318..337
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
promoter 429..484
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 435..453
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 465..483
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 2641..2652
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4079..4097)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4093..4140
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(4370..5185)
/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 5093..5112
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(5186..5277)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5295..5930)
/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(5440..5459)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6190..410)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
ORIGIN
1 acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct
61 aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt
121 gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa
181 tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca
241 atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc
301 tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta
361 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat cattaattcc
421 taatttttgt tgacactcta tcattgatag agttatttta ccactcccta tcagtgatag
481 agaaaagtga actctagaaa taattttgtt taactttaac ctgtagaaat aattttgttt
541 aactttaata aggagatata ccatggcgac aagtttacct acgccctcag caattacgac
601 ttcggcgtta gagtatgcat tccatactcc cgctcgcaat ctaacgaaat ctcgcggaaa
661 aaacattcat cgttatgtca gtgtaaagat gagtaagagg attacggtag aatctactct
721 agagtgtgat gcctgctatc actttgattt tgagccaagt attgttcgct tttgcgctca
781 accgattcga tttttatatt atctcaatgg tcagtctcac tcctatgttc ctgactttct
841 agttcaattt gataccaacg agtttgttct atatgaagta aagtcagctt atgctaagaa
901 caaacctgat tttgatgttg aatgggaggc gaaagtaaaa gcagcaactg aactagggct
961 agaattggag cttgttgaag agagtgatat tagggatacg gttgtattaa ataatcttaa
1021 gcgcatgcat cgttatgctt cgaaagatga gctgaataac gtacataact ctctcttaaa
1081 aataataaag tacaatggcg cccaatctgc aagatgcttg ggagaacagt tgggtttaaa
1141 aggccgaact gttttaccaa ttttgtgcga tttgctgtca aggtgtttac tcgatacacg
1201 tttggataag cctctatctc ttgaatctcg atttgagttg gccagttatg gctaagaaag
1261 ggttctcaag tttccataga aaagcagtct cttcccaaga tacgcttgaa tcgatagagc
1321 ttgtctctag cgctaattgt ctagaaagtg ttacgtatca agatatatca gcatttcccg
1381 aaacaattgc ggtagagatt aatttccgat taagcattct tcgcttttta gcgcgaaagt
1441 gcgaaaccat tgtggccaaa tcaattgaac cacatcgtgt agagctacag caaaactata
1501 gtagaaaaat acccagtgca ataacgatat atcgatggtg gcttgctttt cgaaaatcag
1561 actacaaccc cattagctta gcacctaata tcaaggatag aggtaataga gagacaaaag
1621 tgtcaacagt tgttgattct attatggaac aggcagttga aagagttata tctggacgaa
1681 aagtcaatgt tagctctgca tataaacgtg ttcgacgaaa agttcgtcaa tacaatctaa
1741 ctcatggaac gaaatacacg tatcctaagt acgaatctgt aagaaagcga gtaaaaaaga
1801 aaaccccatt tgagttatta gccgcaggga aaggggagag agtagctaag agagagtttc
1861 gccgaatggg aaaaaagatc ctcacgtcta gcgtgctaga gagggttgaa atagatcaca
1921 ctgtcgttga cctttttgca gtacacgaag agtatcgaat cccattgggc cgaccttggc
1981 ttactcaatt ggttgattgt tacagtaaag ctgtaatcgg tttttattta ggtttcgagc
2041 ctcctagcta tgtgtcggtt tcccttgcac ttaagaatgc aatacaacgc aaagatgact
2101 taatctcctc gtatgaatcg atcgagaatg aatggctatg ttatggcatc ccagacctac
2161 tcgtaactga taatggtaaa gagtttttgt cgaaagcatt tgatcaagca tgtgaatcac
2221 tattgatcaa tgtgcatcaa aataaagttg agacgcccga caacaaacct catgttgaac
2281 gtaactacgg gactattaat acttctctgt tagacgattt acctgggaaa tccttcagcc
2341 agtaccttca aagagaaggg tacgactctg tgggagaagc tacccttaca ctcaatgaga
2401 ttagagaaat ttacttaatt tggttggtgg atatttatca taaaaaaccc aatcagagag
2461 gcactaattg tcctaatgtt gcgtggaaaa agggttgtca agaatgggaa ccagaggagt
2521 tctctggttc taaagacgaa ttagacttta aatttgctat tgttgattac aaacaactta
2581 ctaaagtagg gataactgtc tacaaagaac tgagttatag caatgaccgt ttagctgaat
2641 atagagggaa gaaaggaaac cataaagttc agttcaagta taaccctgag tgtatggcag
2701 ttatttgggt gttggatgag gatatgaatg agtactttac agttaatgcg attgactacg
2761 aatatgcaag tagagtatca ctttggcaac ataaatataa catgaaatat caagcagaac
2821 taaattcagc agaatatgat gaggacaagg aaattgatgc agaaataaaa attgaagaaa
2881 tcgcagatcg ttcaattgtt aagactaaca aaatcagagc tcggaggcgt ggcgctaggc
2941 atcaagagaa tagcgcaagg gctaagtcaa tcagtaatgc gaacccggcc tcgatacaaa
3001 aacatgaaga tgaaatcgtt agtgcagata atgacgattg ggatattgat tatgtctgag
3061 aaatcgtcaa atgagtgaaa cgcgtgaggc tcgtatatca agagctaaaa gggcatttgt
3121 atccacaccg agcgttagga aaatcttgag ttacatggat agatgtagag atctatcaga
3181 cctagagtct gagcctacat gcatgatggt ctatggtgct tcgggtgtag gtaaaacgac
3241 cgtcatcaag aaatacttaa atcaaaacag aagagagtcc gaagccgggg gcgatataat
3301 accggttttg catattgagt tgccagacaa tgcgaagcca gtagatgcag caagggaatt
3361 gctggttgaa atgggtgacc cgctagcact ttatgaaact gacttagcta gattgacgaa
3421 aagactgact gaattaatcc ctgcggtcgg cgtgaagctg attattatcg atgagttcca
3481 acatttggtg gaagaaaggt caaatcgggt tcttacccaa gtaggtaatt ggctaaaaat
3541 gatacttaac aaaacgaaat gtccaattgt tatatttggt atgccatact caaaagttgt
3601 actgcaagca aactcgcaac ttcacgggcg attttccatt caggttgaac tgcgcccctt
3661 tagctaccag ggaggtagag gtgtatttaa aacttttttg gaataccttg ataaagccct
3721 accttttgaa aaacaggctg gcttagccaa cgaaagtttg cagaaaaaat tgtatgcatt
3781 ctctcaggga aacatgcgtt cgttgagaaa ccttatttat caagcatcta tcgaagcaat
3841 tgataatcag catgagacga taaccgaaga agatttcgtt tttgcatcga agttgacatc
3901 gggcgataaa cccaactcat ggaaaaatcc ttttgaggag ggtgttgagg taacagaaga
3961 tatgttacga ccgccaccaa aagatattgg ttgggaggac tatttgagac attcaacccc
4021 gagagtgagt aaaccaggta gaaataaaaa ctttttcgaa taacctaggc tgctgccacc
4081 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
4141 ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
4201 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgacaagc
4261 tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
4321 tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact
4381 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
4441 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
4501 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
4561 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
4621 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
4681 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
4741 gacgaaatac gcggtcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
4801 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
4861 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
4921 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
4981 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
5041 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
5101 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag
5161 acgtttcccg ttgaatatgg ctcatactct tcctttttca atattattga agcatttatc
5221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
5281 gcatgctagc gcagaaacgt cctagaagat gccaggagga tacttagcag agagacaata
5341 aggccggagc gaagccgttt ttccataggc tccgcccccc tgacgaacat cacgaaatct
5401 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
5461 ctgatggctc cctcttgcgc tctcctgttc ccgtcctgcg gcgtccgtgt tgtggtggag
5521 gctttaccca aatcaccacg tcccgttccg tgtagacagt tcgctccaag ctgggctgtg
5581 tgcaagaacc ccccgttcag cccgactgct gcgccttatc cggtaactat catcttgagt
5641 ccaacccgga aagacacgac aaaacgccac tggcagcagc cattggtaac tgagaattag
5701 tggatttaga tatcgagagt cttgaagtgg tggcctaaca gaggctacac tgaaaggaca
5761 gtatttggta tctgcgctcc actaaagcca gttaccaggt taagcagttc cccaactgac
5821 ttaaccttcg atcaaaccgc ctccccaggc ggttttttcg tttacagagc aggagattac
5881 gacgatcgta aaaggatctc aagaagatcc tttacggatt cccgacacca tcactctaga
5941 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
6001 tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
6061 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
6121 gttgcgcctg cctaatgatt tttatcaaaa tcattaaggt taccatcacg gaaaaaggtt
6181 atgctgcttt taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa
6241 ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg
6301 tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg
6361 atgctcttga tcttccaata cgcaacctaa agtaaaatgc ccc
//
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