基本信息

别名:	pBAD-gIIIA
启动子:	araBad
复制子:	PBR322 ori
质粒分类:	大肠杆菌载体；pBAD系列表达质粒
质粒大小:	4145bp
质粒标签:	N-GIII signal, C-His, C-Myc
原核抗性:	氨苄青霉素Amp
克隆菌株:	Top10
培养条件:	37℃，有氧，LB
表达宿主:	推荐LMG194
培养条件:	37℃，有氧，LB
诱导方式:	阿拉伯糖
5'测序引物:	pBAD-F: ATGCCATAGCATTTTTATCC
3'测序引物:	pBAD-R: GATTTAATCTGTATCAGG

质粒简介

pBAD/gIII载体质粒是衍生于pBR322载体。载体设计用来在大肠杆菌中进行可调节分泌性表达和纯化重组目的蛋白。使用基因III信号序列来定位重组蛋白，使其分泌到大肠杆菌周质腔中。使用大肠杆菌araBAD启动子（pBAD）增强了大肠杆菌重组蛋白可溶性表达的水平。载体上的调节蛋白AraC能够调控pBad启动子。

质粒图谱

质粒序列

LOCUS Exported 4145 bp ds-DNA circular SYN 12-SEP-2016
DEFINITION synthetic circular DNA
KEYWORDS pBAD gIII A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4145)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, September 12, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..4145
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..285
/gene="araBAD"
/note="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)"
CDS 427..456
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 472..489
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 715..801
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 893..920
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 940..1031
/gene="bla"
/note="AmpR promoter"
CDS 1032..1892
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2063..2651
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2837..2977
/note="bom"
/note="basis of mobility region from pBR322"
CDS complement(3241..4119)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/note="araC"
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
ORIGIN
1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
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 gaaaaaactg ctgttcgcga ttccgctggt ggtgccgttc
361 tatagccata gcaccatgga gctcgagatc tgcagctggt accatatggg aattcgaagc
421 tttctagaac aaaaactcat ctcagaagag gatctgaata gcgccgtcga ccatcatcat
481 catcatcatt gagtttaaac ggtctccagc ttggctgttt tggcggatga gagaagattt
541 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
601 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
661 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
721 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
781 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
841 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
901 atcctgacgg atggcctttt tgcgtttcta caaactcttt ttgtttattt ttctaaatac
961 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
1021 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
1081 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
1141 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
1201 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
1261 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
1321 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
1381 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
1441 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
1501 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
1561 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
1621 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
1681 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
1741 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
1801 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
1861 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
1921 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
1981 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
2041 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
2101 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
2161 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
2221 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
2281 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
2341 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
2401 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
2461 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
2521 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
2581 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
2641 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
2701 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
2761 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
2821 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
2881 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
2941 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
3001 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
3061 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca
3121 gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc
3181 agggattctg caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa
3241 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
3301 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
3361 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
3421 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
3481 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
3541 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
3601 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
3661 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
3721 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
3781 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
3841 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
3901 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
3961 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
4021 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
4081 gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt
4141 cagag