| 质粒名称: | pBXNH3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 47067 |
| 存储实验室: | Raimund Dutzler, Eric Geertsma |
|
载体骨架基本信息 |
|
| 载体名称: | pBAD24 |
| 载体出品公司: | LM Guzman |
| 载体抗性: | 氯霉素和氨苄青霉素 |
| 空载体大小: | 5779 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 与FX克隆和添加的标签/融合蛋白兼容 |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | araBAD |
| 标签/融合蛋白: |
decaHis (10x His)（主干上的N终端）
3C protease site (PreScission site) （主干上的N端）
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | pBAD-F |
| 3' 测序引物 | pBAD-R |
|
文献引用方法 |
|
| 材料和方法部分: | pBXNH3 was a gift from Raimund Dutzler & Eric Geertsma (Addgene plasmid # 47067 ; http://n2t.net/addgene:47067 ; RRID:Addgene_47067) |
| 参考文献部分: | A versatile and efficient high-throughput cloning tool for structural biology. Geertsma ER, Dutzler R. Biochemistry. 2011 Apr 19;50(15):3272-8. Epub 2011 Mar 25. 10.1021/bi200178z PubMed 21410291 |
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 pBXNH3 5779 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION FX cloning E.coli expression vector with araBAD promoter and
N-terminal 10x His tag and 3C protease cleavage site.
ACCESSION .
VERSION .
KEYWORDS pBXNH3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5779)
AUTHORS Geertsma ER, Dutzler R
TITLE A versatile and efficient high-throughput cloning tool for
structural biology.
JOURNAL Biochemistry. 2011 Apr 19;50(15):3272-8. Epub 2011 Mar 25.
PUBMED 21410291
REFERENCE 2 (bases 1 to 5779)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5779
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..285
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 208..227
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 322..351
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 352..375
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS complement(424..729)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(526..545)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS complement(1071..1730)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1645..1664
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(1784..1814)
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind complement(1839..1963)
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS complement(1982..2002)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(2006..2026)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2062..2091
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2107..2124
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2180..2197)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 2350..2436
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2528..2555
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2574..2665
/gene="bla"
/label=AmpR promoter
CDS 2666..3526
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2884..2903)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3697..4285
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4186..4205
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4439..4456
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4471..4611
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4537..4556)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4697..4719
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4875..5753)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggcact
61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
301 taacaggagg aattaaccat gcaccatcat catcaccatc atcatcatca tttagaagtt
361 ttgtttcaag gtccaagtag aagagcgagc tgcagactgg ctgtgtataa gggagcctga
421 catttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct
481 gagatcagcc acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat
541 catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg agactttatc
601 tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat
661 atcactctgt acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt
721 aaactgcatt tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc
781 gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac
841 gggcttcatt ctgcatggtt gtgcttacca gaccggagat attgacatca tatatgcctt
901 gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct
961 ctttttgaca tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg
1021 cgcaaatacg catactgtta tctggctttt agtaagccgg atccacgcgt ttacgccccg
1081 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
1141 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
1201 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
1261 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
1321 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
1381 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
1441 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
1501 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
1561 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
1621 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
1681 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
1741 ttagctcctg aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa
1801 gcataaagtg taaagcctgg ggtgcctaat gcggccgcca tagtgactgg atatgttgtg
1861 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
1921 tatcatttta cgtttctcgt tcagcttttt tgtacaaact tgtgatatcg aattcgctag
1981 ctacctttct cttctttttt ggatctacct ttctcttctt ttttggggat ccactagttc
2041 agctcttctg cataataact agaacaaaaa ctcatctcag aagaggatct gaatagcgcc
2101 gtcgaccatc atcatcatca tcattgagtt taaacggtct ccagcttggc tgttttggcg
2161 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
2221 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
2281 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
2341 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
2401 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
2461 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
2521 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
2581 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
2641 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
2701 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
2761 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
2821 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
2881 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
2941 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
3001 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
3061 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
3121 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
3181 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
3241 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
3301 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
3361 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
3421 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
3481 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
3541 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
3601 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
3661 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
3721 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
3781 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
3841 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
3901 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
3961 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
4021 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
4081 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
4141 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
4201 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
4261 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
4321 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
4381 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
4441 cgagtcagtg agcgaggaag cggaagtgcg cctgatgcgg tattttctcc ttacgcatct
4501 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
4561 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
4621 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
4681 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
4741 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt
4801 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
4861 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
4921 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
4981 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
5041 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
5101 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
5161 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
5221 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
5281 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
5341 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
5401 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
5461 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
5521 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
5581 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
5641 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
5701 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
5761 atactcccgc cattcagag
//
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