pHB201

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  • Model: PVT5013
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pHB201

PVT5013      Packing 2ug

 

pHB201 Information

Promoter: T7, T3

Replicon: ori

Terminator: T1

Plasmid classification: the wide host series, Bacillus subtilis carrier

Plasmid size: 6594bp

Prokaryotic resistance: Chl

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic LB

Expression host: Bacillus subtilis

Induction mode: IPTG induction

5'sequencing primers: T7:TAATACGACTCACTATAGGG

3'sequencing primers: primers designed according to sequence

Use: Bacillus subtilis

 

pHB201 Description

A genomic library of Bacillus lyticus was constructed in lambda GEM 11 vector and screened for the xylanase gene using Congo red plate assay. A 16-kb fragment containing the xylanase gene was obtained which was further subcloned using Mbo I partial digestion in an E. coli pUC 19 vector. A 1.3-kb sub-fragment was obtained which coded for a xylanase gene of Mr 23,650 Da. This fragment was sequenced and the homology was checked with known xylanases. The maximum homology was 97%, which was obtained with an endo xylanase gene from Bacillus species at the DNA level, while the translated sequence showed only one amino acid change from alanine to serine at position number 102. Expression was checked in E. coli, using the native promoter, and an extracellular activity of 5.25 U/mL was obtained. Cloning of the gene was done in Bacillus subtilis using a shuttle vector pHB 201, which resulted in increasing the basal level xylanase activity from 14.02 to 22.01 U/mL.

pHB201-polyclonal-site

 

pHB201 Sequence

LOCUS       Exported File           6594 bp ds-DNA    circular SYN 29-5-2015

DEFINITION  .

ACCESSION   .

VERSION     .

KEYWORDS    pHB201

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6594)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported 2015-5-29

FEATURES             Location/Qualifiers

     source          1..6594

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        1533..1551

                     /note="T3 promoter"

                     /note="promoter for bacteriophage T3 RNA polymerase"

     misc_feature    1564..1671

                     /note="MCS"

                     /note="pBluescript multiple cloning site"

     primer_bind     1588..1604

                     /note="SK primer"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     primer_bind     complement(1638..1654)

                     /note="KS primer"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     promoter        complement(1680..1698)

                     /note="T7 promoter"

                     /note="promoter for bacteriophage T7 RNA polymerase"

     primer_bind     complement(1711..1727)

                     /note="M13 fwd"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     terminator      2073..2159

                     /gene="Escherichia coli rrnB"

                     /note="rrnB T1 terminator"

                     /note="transcription terminator T1 from the E. coli rrnB 

                     gene"

     terminator      2251..2278

                     /note="rrnB T2 terminator"

                     /note="transcription terminator T2 from the E. coli rrnB 

                     gene"

     CDS             complement(2717..3367)

                     /codon_start=1

                     /note="CamR"

                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE

                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV

                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL

                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"

     rep_origin      6329..323

                     /direction=RIGHT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

ORIGIN

        1 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg

       61 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc

      121 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa

      181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc

      241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt

      301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct

      361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc

      421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg

      481 agtcagtgag cgaggaagcg gaagaagctt actttattgt tttgccattt taaaaggttc

      541 aaataaaata tttttcataa aattatcaga tgttttttaa aaaatgaaat aaaagaagaa

      601 aaaaatagaa aatacactaa cccaaagaag cgcgtaatat cgagggttta agagacaata

      661 agaaaaaaag attgaaaaat gacattaaat tcttgacagg gagagatagg tttgatagaa

      721 tataatagtt gtcgcgagag acgacgttga agcgcgacaa gctagaccat tgaaaactga

      781 ataaagaaga atgactcatg tgatgtagca atacatcaaa aatctgtcaa tcaataatga

      841 aagacaagcc atcaattcgg tgacttaaat actttatttg agagtttgat cctctagagt

      901 cgacctgcag gctttaacgt aggcaaagct cagggtagac tttgaatgga cagaaacatg

      961 acatatctct tgaaaggatg attgtggtgg tgaaaacaga taaaatctcc tcctgaatac

     1021 agtaaatcac attcaggagg agataaaatt gtttaaacaa atagacgaaa attatctgcg

     1081 aaaagagcac tttcaccatt atatgacgtt aacccgatgc tcatatagct tggtgatcaa

     1141 tctagacatc acgaaattgc atgcaatatt aaaagaaaaa aagctgaaag tatatcctgt

     1201 gcaaatttat ttgttagcaa gagctgtgca aaaaattcct gagtttcgga tggatcaagt

     1261 gaacgatgaa cttggttact gggagattct ccatcctagt tatacgattc taaataaaga

     1321 aacaaagacg ttttcaagta tttggacgcc ttttgatgaa aactttgctc agttttataa

     1381 aagctgtgta gccgatattg aaacatttag caaaagcagc aacctatttc cgaaacctca

     1441 tatgccagaa aacatgttca atatttcaag tctaccgtgg attgatttta cttcttttaa

     1501 ccttaatgta tctacagatg aagctagcgc gcaattaacc ctcactaaag ggaacaaaag

     1561 ctggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg gctgcaggaa

     1621 ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccaattcgcc

     1681 ctatagtgag tcgtattacg cgcgcaattc actggccgtc gttttacaac gtcgtgactg

     1741 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg

     1801 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatta

     1861 gagatcaatt ccctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc

     1921 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc

     1981 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc

     2041 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag

     2101 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc

     2161 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc

     2221 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg

     2281 cgtttctaca aactcttcct gtcgtcatat ctacaagcca tccccccaca gatacggtaa

     2341 actagcctcg tttttgcatc aggaaagcag ggaattgatc tgatgggtag aaccttgggc

     2401 tttgcccaaa cccaccaacg gatggccgtt ggattccaac attcaatccc cccaaccccc

     2461 cgcatctccg gggggctccc tcgccggttg gcgtcaaaca aacgagtttg tttgagtgct

     2521 gaaagggttc gggtagcact gagcgagtca actccttaat cctccctacg gtccggttac

     2581 cgttgactca cgccataccc atcgaatggc gaatggcgga tctggagctg taatataaaa

     2641 accttcttca actaacgggg caggttagtg acattagaaa accgactgta aaaagtacag

     2701 tcggcattat ctcatattat aaaagccagt cattaggcct atctgacaat tcctgaatag

     2761 agttcataaa caatcctgca tgataaccat cacaaacaga atgatgtacc tgtaaagata

     2821 gcggtaaata tattgaatta cctttattaa tgaattttcc tgctgtaata atgggtagaa

     2881 ggtaattact attattattg atatttaagt taaacccagt aaatgaagtc catggaataa

     2941 tagaaagaga aaaagcattt tcaggtatag gtgttttggg aaacaatttc cccgaaccat

     3001 tatatttctc tacatcagaa aggtataaat cataaaactc tttgaagtca ttctttacag

     3061 gagtccaaat accagagaat gttttagata caccatcaaa aattgtataa agtggctcta

     3121 acttatccca ataacctaac tctccgtcgc tattgtaacc agttctaaaa gctgtatttg

     3181 agtttatcac ccttgtcact aagaaaataa atgcagggta aaatttatat ccttcttgtt

     3241 ttatgtttcg gtataaaaca ctaatatcaa tttctgtggt tatactaaaa gtcgtttgtt

     3301 ggttcaaata atgattaaat atctcttttc tcttccaatt gtctaaatca attttattaa

     3361 agttcatttg atatgcctcc taaattttta tctaaagtga atttaggagg cttacttgtc

     3421 tgctttcttc attagaatca atcctttttt aaaagtcaat attactgtaa cataaatata

     3481 tattttaaaa atatcccact ttatccaatt ttcgtttgtt gaactaatgg gtgctttagt

     3541 tgaagaataa agaccacatt aaaaaatgtg gtcttttgtg tttttttaaa ggatttgagc

     3601 gtagcgaaaa atccttttct ttcttatctt gataataagg gtaactattg ccgcgcctga

     3661 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataagc cggtcttttt

     3721 cgattttggc gaagccgaaa tcgggtcttt tcttatcttg atactatata gaaacaacat

     3781 catttttcaa aattaggtca aagccttgtg tatcaagggt ttgatggttc tttgacaggt

     3841 aaaaactcct tctgctatta ttaaggtgtc gaatcaaaat aatagaatgc tagagaacta

     3901 gctcagaagg agtttttttg ttgatttatt catctgaaaa tgattatagc atcctcgaag

     3961 ataaaaccgc aacaggtaaa aagcgggatt ggaaggggaa aaagagacgg acgaacctca

     4021 tggcggagca ttacgaagcg ttagagagta agattggggc accttactat ggcaaaaagg

     4081 ctgaaaaact aattagttgt gcagagtatc tttcgtttaa gagagacccg gagacgggca

     4141 agttaaaact gtatcaagcc catttttgta aagtgaggtt atgtccgatg tgtgcgtggc

     4201 gcaggtcgtt aaaaattgct tatcacaata agttgatcgt agaggaagcc aatagacagt

     4261 acggctgcgg atggattttt ctcacgctga cgattcgaaa tgtaaaggga gaacggctga

     4321 agccacaaat ttctgcgatg atggaaggct ttaggaaact gttccagtac aaaaaagtaa

     4381 aaacttcggt tcttggattt ttcagagctt tagagattac caaaaatcat gaagaagata

     4441 catatcatcc tcattttcat gtgttgatac cagtaaggaa aaattatttt gggaaaaact

     4501 atattaagca ggcggagtgg acgagccttt ggaaaaaggc gatgaaattg gattacactc

     4561 caattgtcga tattcgtcga gtgaaaggta aagctaagat tgacgctgaa cagattgaaa

     4621 acgatgtgcg gaacgcaatg atggagcaaa aagctgttct cgaaatctct aaatatccgg

     4681 ttaaggatac ggatgttgtg cgcggtaata aggtgactga agacaatctg aacacggtgc

     4741 tttacttgga tgatgcgttg gcagctcgaa ggttaattgg atacggtggc attttgaagg

     4801 agatacataa agagctgaat cttggtgatg cggaggacgg cgatctggtc aagattgagg

     4861 aagaagatga cgaggttgca aatggtgcat ttgaggttat ggcttattgg catcctggca

     4921 ttaaaaatta cataatcaaa taaaaaaagc agacctttag aaggcctgct tttttaacta

     4981 acccatttgt attgtgttga aatatgtttt gtatggtgca ctctcagtac aatctgctct

     5041 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg

     5101 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg

     5161 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc

     5221 ctatttttat aggttaatgt catgataata atggtttctt agcgattcac aaaaaatagg

     5281 cacacgaaaa acaagttaag ggatgcagtt tatgcatccc ttaacttact tattaaataa

     5341 tttatagcta ttgaaaagag ataagaattg ttcaaagcta atattgttta aatcgtcaat

     5401 tcctgcatgt tttaaggaat tgttaaattg attttttgta aatattttct tgtattcttt

     5461 gttaacccat ttcataacga aataattata cttttgttta tctttgtgtg atattcttga

     5521 tttttttcta cttaatctga taagtgagct attcacttta ggtttaggat gaaaatattc

     5581 tcttggaacc atacttaata tagaaatatc aacttctgcc attaaaagta atgccaatga

     5641 gcgttttgta tttaataatc ttttagcaaa cccgtattcc acgattaaat aaatctcatt

     5701 agctatacta tcaaaaacaa ttttgcgtat tatatccgta cttatgttat aaggtatatt

     5761 accatatatt ttataggatt ggtttttagg aaatttaaac tgcaatatat ccttgtttaa

     5821 aacttggaaa ttatcgtgat caacaagttt attttctgta gttttgcata atttatggtc

     5881 tatttcaatg gcagttacga aattacacct ctttactaat tcaagggtaa aatggccttt

     5941 tcctgagccg atttcaaaga tattatcatg ttcatttaat cttatatttg tcattatttt

     6001 atctatatta tgttttgaag taataaagtt ttgactgtgt tttatatttt tctcgttcat

     6061 tataaccctc tttaatttgg ttatatgaat tttgcttatt aacgattcat tataaccact

     6121 tattttttgt ttggttgata atgaactgtg ctgattacaa aaatactaaa aatgcccata

     6181 ttttttcctc cttataaaat tagtataatt atagcacgaa aaggatctag gtgaagatcc

     6241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag

     6301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct

     6361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac

     6421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc

     6481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg

     6541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt ctta

//
 

Caution:
Product is for research use only!

 

Search name
pHB201,Plasmid pHB201,pHB201 vector

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