| 质粒名称: | pATetO 6XHis-Taq DNA Ligase |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 178798 |
| 存储实验室: | Andrew Ellington |
|
载体骨架基本信息 |
|
| 载体名称: | pATetO |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 4869 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Taq DNA连接酶 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CTCTCCCTATCAGTGATAGAGAAAACTAG |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pATetO 6XHis-Taq DNA Ligase was a gift from Andrew Ellington (Addgene plasmid # 178798 ; http://n2t.net/addgene:178798 ; RRID:Addgene_178798) |
| 参考文献部分: | Producing molecular biology reagents without purification. Bhadra S, Nguyen V, Torres JA, Kar S, Fadanka S, Gandini C, Akligoh H, Paik I, Maranhao AC, Molloy J, Ellington AD. PLoS One. 2021 Jun 1;16(6):e0252507. doi: 10.1371/journal.pone.0252507. eCollection 2021. 10.1371/journal.pone.0252507 PubMed 34061896 |
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 pATetO 6XHis-Taq DNA Ligase 4869 bp ds-DNA circular SYN 07-MAR-2022
DEFINITION Inducible expression of Taq DNA Ligase in E. coli.
ACCESSION .
VERSION .
KEYWORDS pATetO 6XHis-Taq DNA Ligase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4869)
AUTHORS Bhadra S, Nguyen V, Torres JA, Kar S, Fadanka S, Gandini C, Akligoh
H, Paik I, Maranhao AC, Molloy J, Ellington AD
TITLE Producing molecular biology reagents without purification.
JOURNAL PLoS One. 2021 Jun 1;16(6):e0252507. doi:
10.1371/journal.pone.0252507. eCollection 2021.
PUBMED 34061896
REFERENCE 2 (bases 1 to 4869)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 7, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4869
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(138..764)
/codon_start=1
/gene="tetR"
/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="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRDNAKSFRCALLSHRDGAKVHLGT
RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 669..688
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
CDS complement(771..1631)
/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 1394..1413
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1632..1736)
/gene="bla"
/label=AmpR promoter
promoter 1889..1944
/gene="tetR"
/label=tetR/tetA promoters
/note="overlapping promoters for bacterial tetR and tetA"
protein_bind 1895..1913
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 1925..1943
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 1958..1980
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2006..2023
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
rep_origin complement(4266..4854)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4346..4365)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
61 agattatcaa aaaggatctt cacctagatc cttttaaact agtgaagtta ccatcacgga
121 aaaaggttat gctgctttta agacccactt tcacatttaa gttgtttttc taatccgcat
181 atgatcaatt caaggccgaa taagaaggct ggctctgcac cttggtgatc aaataattcg
241 atagcttgtc gtaataatgg cggcatacta tcagtagtag gtgtttccct ttcttcttta
301 gcgacttgat gctcttgatc ttccaatacg caacctaaag taaaatgccc cactgcgctg
361 agtgcatata atgcattctc tagtgaaaaa ccttgttggc ataaaaaggc taattgattt
421 tcgagagttt catactgttt ttctgtaggc cgtgtaccta aatgtacttt tgctccatcg
481 cgatgactta gtaaagcaca tctaaaactt ttagcgttat cacgtaaaaa atcttgccag
541 ctttcccctt ctaaagggca aaagtgagta tggtgcctat ctaacatctc aatggctaag
601 gcgtcgagca aagcccgctt attttttaca tgccaataca atgtaggctg ctctacacct
661 agcttctggg cgagtttacg ggttgttaaa ccttcgattc cgacctcatt aagcagctct
721 aatgcgctgt taatcacttt acttttatct aaacgagaca tcattaattc ctaccaatgc
781 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
841 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
901 atgataccgc gagagccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
961 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctatcaat
1021 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
1081 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
1141 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
1201 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
1261 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
1321 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
1381 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
1441 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
1501 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
1561 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
1621 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
1681 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
1741 tttcccggga actgccagac atcaaataaa acaaaaggct cagtcggaag actgggcctt
1801 ttgttttatc tgttgtttgt cggtgaacac tctcccgatt gcgatacaga ccctaatttc
1861 acatcatatg acactaatcc taatttttgt tgacactcta tcattgatag agttatttta
1921 cctctcccta tcagtgatag agaaaactag aaataatttt gtttaacttt aagaaggaga
1981 tatacatatg gctagcagag gatcgcatca ccatcaccat cacatgaagg gcgcatgacc
2041 ctggaggccg cgcgccgtcg cgtgaacgag ttacgcgatt taattcgtta tcacaactat
2101 ctttactacg tgttggacgc ccctgaaatc tcagatgcag agtatgatcg cctgttacgc
2161 gaacttaagg agctggaaga acgtttccct gagttacaga gccctgatag cccgactgaa
2221 caagtagggg cacgcccgct tgaatccact ttccgccctg tgcgtcatcc gacacgtatg
2281 tattctttag acaatgcgtt cagtttggat gaagtacgtg cgttcgaaga gcgcattgag
2341 cgcgcgttgg gtcgtaaggg gcctttttta tataccgtgg agcataaggt tgatgggtta
2401 tcagttaatt tgtattacga agaaggtatc ttagtatttg gcgcaacgcg cggcgatggg
2461 gaaacgggtg aagaagtgac acaaaacctt cttactattc gcactatccc gcgtcgtctt
2521 acgggagtac ctgatcgcct tgaagttcgt ggagaagtat acatgcccat cgaggcattt
2581 cttcgtttaa accaagagtt agaagaagca ggcgaacgca ttttcaagaa cccgcgcaat
2641 gccgcagccg gttcgcttcg tcaaaaggac ccccgtgtta ctgctcgtcg cggattgcgc
2701 gcgacgttct acgccttggg gttgggtctt gaagagacgg ggcttaaatc gcagcatgac
2761 cttttgttgt ggcttcgtga gcgtggtttc ccagtagaac acggatttac acgtgctttg
2821 ggggcggaag gggtggagga agtttaccaa gcctggttaa aggagcgccg taagcttccc
2881 ttcgaagcgg acggtgttgt cgtcaaattg gatgacctgg cactgtggcg tgagttgggg
2941 tatacggcac gtgcaccacg tttcgcgtta gcctataagt ttcccgcaga ggagaaggag
3001 acgcgtctgt tatcagtcgc tttccaagta ggacgtaccg ggcgtattac tccagtggga
3061 gtacttgaac ccgtttttat cgaggggtca gaggtgtcgc gtgttacgtt acataacgag
3121 agtttcattg aagagttgga tgtacgtatt ggggattggg tacttgtcca caaagctggc
3181 ggcgtgatcc ctgaggtctt gcgcgtcctg aaagaacgcc gcaccgggga ggagaaacca
3241 attttgtggc ctgaaaattg tcctgaatgt gggcatgccc tgatcaagga agggaaagta
3301 catcgttgcc ctaacccact gtgccccgct aagcgtttcg aagcgattcg tcattacgcc
3361 agccgcaaag ctatggatat ccaaggttta ggggagaaat taattgaaaa actgctggag
3421 aaaggcttag tgcgtgatgt ggctgattta taccgtctta aaaaggagga cttggtaaac
3481 ttggaacgta tgggcgagaa aagtgcggag aatttgttac gccagatcga ggagtccaaa
3541 gggcgcgggc tggaacgcct gttatacgcc ttggggctgc ccggcgtggg tgaagtgctt
3601 gcgcgtaatt tagccttacg cttcggccac atggaccgtc ttttagaggc cgggcttgag
3661 gaccttcttg aggtggaagg tgtcggcgag ttgactgctc gcgcgattct gaatactctg
3721 aaagaccctg agtttcgtga tttagttcgc cgtttgaaag aagcgggtgt cgaaatggaa
3781 gctaaagaac gcgaaggtga agcattgaag ggtttgacgt tcgtcattac gggtgaactg
3841 agccgcccgc gtgaggaagt caaagctctt ttgcgtcgct taggcgcaaa agtgaccgac
3901 tccgtctctc gcaaaaccgg ttttttggta gtaggcgaga acccggggtc gaagttagaa
3961 aaggcacgtg ctttaggagt ccccacactg tccgaagaag agctgtatcg ccttatcgag
4021 gagcgtacag ggaaagatcc ccgtgcgctg acggcatgat aattgcctct gccaaaattc
4081 tgtcctcaag cgttttagtt cgtcggtcag tttcacctga tttacgtaaa aacccgcttc
4141 ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtcggccat tcgatggtgt
4201 cgggtcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
4681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
4861 cctttgatc
//
