pAX01 Plasmid

pAX01

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pAX01,Plasmid pAX01,pAX01 vector

pAX01 Information

Promoter: xylA promoter

Replicon: ColE1 ori

Terminator: T1 terminator, T2 Terminator

Plasmid classification: hay series plasmid, hay integration plasmid, plasmid in hay.

Plasmid size: 7781bp

Prokaryotic resistance: ampicillin Amp (100 u g/ml)

Screening markers: erythromycin ERM

Cloned strains of Escherichia coli, DH5 A and other Escherichia coli

Culture conditions: 37 centigrade, aerobic LB

Expression host: Bacillus subtilis, BS168 and other Bacillus subtilis

Inducement: xylose induction

5'sequencing primers: pAX-F (GTTGCCCTGGAGACAGGGG)

3'sequencing primers: pAX-R (GATATGGTGCAAGTCAGCACG)

Use: Bacillus subtilis

pAX01 Description

pAX01 plasmid has no pathogenicity, safety and no obvious codon bias. Adding signal peptide can directly secrete the expressed protein into the medium (at present, about 60% of the commercially available enzymes are produced by Bacillus subtilis), the expression product is soluble, the expression product is separated from the intracellular protein, and it does not need to break the cells, which is beneficial to the separation and purification, and is beneficial to the separation and purification. The mechanism of Bacillus subtilis transcription, translation, protein folding and secretion is clear, which is conducive to genetic manipulation and large-scale fermentation. Based on Escherichia coli bacillus subtilis shuttle plasmid expression vector pAX01, high expression of recombinant foreign protein in Bacillus subtilis can be achieved. The full length of the pAX01 plasmid is 7781bp, and the carrier is based on the groE operon of the Bacillus subtilis based on strong Sigma A- dependent promoter, and is transformed into a highly efficient and controllable promoter by adding operon.

pAX01 expression vector is a Bacillus subtilis, replicon plasmid ori, size 7781bp, e. (Amp); Bacillus subtilis (erythromycin resistance), can be transformed into Escherichia coli DH5 alpha, the coliform (Amp); Bacillus subtilis (erythromycin resistance) plate culture screening a large number of replication amplification culture conditions is LB, 37 C.

pAX01 Sequence

  LOCUS       Exported File           7781 bp ds-DNA    circular SYN 18-4-2015    KEYWORDS    pAX01    SOURCE      synthetic DNA construct      ORGANISM  synthetic DNA construct    REFERENCE   1  (bases 1 to 7781)      TITLE     Direct Submission      JOURNAL   Exported 2015-4-18      FEATURES             Location/Qualifiers         source          1..7781                         /organism="synthetic DNA construct"                         /mol_type="other DNA"         terminator      660..687                         /note="rrnB T2 terminator"                         /note="transcription terminator T2 from the E. coli rrnB                          gene"         terminator      706..792                         /gene="Escherichia coli rrnB"                         /note="rrnB T1 terminator"                         /note="transcription terminator T1 from the E. coli rrnB                          gene"         promoter        2078..2182                         /gene="bla"                         /note="AmpR promoter"         promoter        4352..4456                         /gene="bla"                         /note="AmpR promoter"         CDS             4457..5317                         /codon_start=1                         /gene="bla"                         /product="beta-lactamase"                         /note="AmpR"                         /note="confers resistance to ampicillin, carbenicillin, and                         related antibiotics"                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI                         ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS                         PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW                         EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA                         LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS                         LIKHW"         rep_origin      5488..6076                         /direction=RIGHT                         /note="ori"                         /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of                          replication"         misc_feature    6262..6402                         /note="bom"                         /note="basis of mobility region from pBR322"         CDS             complement(6504..6695)                         /codon_start=1                         /gene="rop"                         /product="Rop protein, which maintains plasmids at low copy                         number"                         /note="rop"                         /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA                         DELYRSCLARFGDDGENL"    ORIGIN            1 ggggtgatgt caaagcttga aaaaacgcac gtaacaaaag caaaatttat gctccatggg           61 ggagactaca accccgatca gtggctggat cggcccgata ttttagctga cgatatcaaa          121 ctgatgaagc tttctcatac gaatacgttt tctgtcggca tttttgcatg gagcgcactt          181 gagccggagg agggcgtata tcaatttgaa tggctggatg atatttttga gcggattcac          241 agtataggcg gccgggtcat attagcaacg ccgagcggag cccgtccggc ctggctgtcg          301 caaacctatc cggaagtttt gcgcgtcaat gcctcccgcg tcaaacagct gcacggcgga          361 aggcacaacc actgcctcac atctaaagtc taccgagaaa aaacacggca catcaaccgc          421 ttattagcag aacgatacgg acatcacccg gcgctgttaa tgtggcacat ttcaaacgaa          481 tacgggggag attgccactg tgaatcgatg cggccgctct agagcaacgt tcttgccatt          541 gctgcataaa aaacgcccgg cggcaaccga gcgttctgaa ttaattaatc atcgggaaga          601 tcttcatcac cgaaacgcgg caggcagctc tagagttaac aagagtttgt agaaacgcaa          661 aaaggccatc cgtcaggatg gccttctgct tagctagagc ggcggatttg tcctactcag          721 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt          781 tcgttttatt tgatgcctca agctagagag tcattaccag atctcactgc agagatcccc          841 gggtctagag tctagggacc tctttagctc cttggaagct gtcagtagta tacctaataa          901 tttatctaca ttccctttag taacgtgtaa ctttccaaat ttacaaaagc gactcataga          961 attatttcct cccgttaaat aatagataac tattaaaaat agacaatact tgctcataag         1021 taacggtact taaattgttt actttggcgt gtttcattgc ttgatgaaac tgatttttag         1081 taaacagttg acgatattct cgattgaccc attttgaaac aaagtacgta tatagcttcc         1141 aatatttatc tggaacatct gtggtatggc gggtaagttt tattaagaca ctgtttactt         1201 ttggtttagg atgaaagcat tccgctggca gcttaagcaa ttgctgaatc gagacttgag         1261 tgtgcaagag caaccctagt gttcggtgaa tatccaaggt acgcttgtag aatccttctt         1321 caacaatcag atagatgtca gacgcatggc tttcaaaaac cactttttta ataatttgtg         1381 tgcttaaatg gtaaggaata ctcccaacaa ttttatacct ctgtttgtta gggaattgaa         1441 actgtagaat atcttggtga attaaagtga cacgagtatt cagttttaat ttttctgacg         1501 ataagttgaa tagatgactg tctaattcaa tagacgttac ctgtttactt attttagcca         1561 gtttcgtcgt taaatgccct ttacctgttc caatttcgta aacggtatcg gtttctttta         1621 aattcaattg ttttattatt tggttgagta ctttttcact cgttaaaaag ttttgagaat         1681 attttatatt tttgttcatg taatcactcc ttcttaatta caaattttta gcatctaatt         1741 taacttcaat tcctattata caaaatttta agatactgca ctatcaacac actcttaagt         1801 ttgcttctaa gtcttatttc cataacttct tttacgtttc cgccattctt tgctgtttcg         1861 atttttatga tatggtgcaa gtcagcacga acacgaaccg tcttatctcc cattatatct         1921 ttttttgcac tgattggtgt atcatttcgt ttttcttttg tgctagagga tcaattcttg         1981 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt         2041 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt         2101 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca         2161 ataatattga aaaaggaaga gtgcggccgc ccgcgggagc tcggatccca tttccccctt         2221 tgatttttag atatcactag tttggaccat ttgtcatttc cccctttgat ttaagtgaac         2281 aagtttatcc atcaactatc ttaattgagt tagtttgttt atccaataaa ctaactttat         2341 ctcatcatat acaaaataaa tgtttatttc aatgtttttt ttagaaaatt tagttataat         2401 attagatatg atacttttaa atatctaatt caagcttcaa aaaacaccaa cttagttcgg         2461 tggataaaca aaggagtggt tattattcaa attgcagatc aagctttagt aaaaaaaatg         2521 aatcaaaaat taatattaga tgaaattttg aagaactccc ctgtctccag ggcaactctc         2581 tctgagatta caggattaaa caagtctact gtctcctctc aagtaaatac actgcttgaa         2641 aaagatttta tttttgaaat tggggcaggg caatctagag gcggcagaag acctgtaatg         2701 cttgttttta ataagaatgc aggctactcg attggtattg atataggagt cgactatctt         2761 aacggaattc taaccgactt agaaggaaat attattctcg agaagacttc tgacttgtct         2821 agttcttccg ctagtgaagt aaaagagatt ttatttgcac ttattcatgg ttttgtaacc         2881 catatgcctg agtcccctta tggtctagtc ggaataggaa tttgtgttcc aggccttgta         2941 gatcgtcatc agcaaattat tttcatgcct aacttaaatt ggaatatcaa agatttgcag         3001 tttttaattg agagtgagtt taatgttccg gtttttgttg aaaatgaagc taatgcagga         3061 gcatacggtg aaaaagtatt tggtatgaca aaaaactatg aaaacatcgt ttacatcagt         3121 attaatatcg gaattggaac tggacttgtt attaacaacg aattgtataa aggtgttcag         3181 ggtttttctg gggaaatggg tcatatgacg atagatttta atggacccaa atgcagctgt         3241 ggaaatcgag gctgttggga attatatgct tctgaaaaag cgttactggc ttcgctctct         3301 aaagaagaaa agaatatttc tcgaaaagag attgtggaac gcgcaaataa aaatgatgta         3361 gaaatgttaa atgcacttca aaactttggc ttttatatcg gaattggatt aaccaatatc         3421 cttaatacat ttgatataga agctgttatc ttgagaaatc atataattga atctcatccc         3481 attgttttaa atacgattaa aaacgaagtt tcttctagag tccattctca tttagacaat         3541 aaatgtgaac tattgccttc ttcgttagga aaaaatgcac ctgctttagg agcggtttct         3601 atcgttattg attctttttt aagtgttacc cctataagtt aggagctccc cgggacgttc         3661 ttgccattgc tgcataaaaa acgcccggcg gcaaccgagc gttctgaatt aattaatcat         3721 cgcgactgca gagatatcga tttcaagcta tatttggagt tgagcctctt gaaacggaca         3781 ccctgtatcc gaaggatcga aacgctgtca gctaccgcag ccaaatatat gaaatgaagg         3841 attatgcaac cgtgattgat gtaaagacag cttcagtgga agcggtgtat caagaagatt         3901 tttatgcgcg cacgccagcg gtcacaagcc atgagtatca gcagggcaag gcgtatttta         3961 tcggcgcgcg tttggaggat caatttcagc gtgatttcta tgagggtctg atcacagacc         4021 tgtctctctc tccagttttt ccggttcggc acggaaaagg cgtctccgta caagcgaggc         4081 aggatcagga caatgattat atttttgtca tgaatttcac ggaagaaaaa cagctggtca         4141 cgtttgatca gagtgtgaag gacataatga caggagacat attgtcaggc gacctgacga         4201 tggaaaagta tgaagtgaga attgtcgtaa acacacatta gcccatcatt cttgaagacg         4261 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta         4321 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta         4381 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata         4441 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc         4501 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga         4561 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct         4621 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg         4681 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta         4741 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat         4801 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt         4861 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga         4921 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga         4981 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga         5041 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc         5101 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc         5161 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg         5221 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat         5281 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata         5341 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct         5401 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga         5461 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg         5521 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc         5581 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct         5641 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc         5701 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt         5761 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg         5821 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca         5881 ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag         5941 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag         6001 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg         6061 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg         6121 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac         6181 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt         6241 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat         6301 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca         6361 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca         6421 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg         6481 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg         6541 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca         6601 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg         6661 gccatgttaa gggcggtttt ttcctgtttg gtcacttgat gcctccgtgt aagggggaat         6721 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta         6781 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga         6841 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg         6901 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc         6961 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg         7021 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg         7081 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca         7141 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc         7201 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca         7261 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt         7321 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg         7381 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc         7441 cgaggcggca taaatcgccg tgacgatcag cggtccagtg atcgaagtta ggctggtaag         7501 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat         7561 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc         7621 catccagcct cgcgtcgcga ctaagaaaat gccgtcaaat ccgctcgcca tgacttcact         7681 aacgatgcct ttgaaaatct tcaagttctt ttctactaat tcaaggcgtg tctcaccagg         7741 tttttggttt gctccggcgc aaatgcagac aatatcagga t   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