pBE2-R

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  • Model: PVT5016
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pBE2-R

PVT5016    2ug

 

pBE2-R Informaiton

 

Promoter: P43 promoter

Replicon: ori

Plasmid classification: Bacillus subtilis plasmid; free plasmid of Bacillus subtilis; secretory plasmid of Bacillus subtilis

Plasmid size: 7014bp

Prokaryotic resistance: ampicillin Amp

Cloning strain: Escherichia coli Stbl3

Culture conditions: 37 C, aerobic, LB

Expression host: Bacillus subtilis

5'Sequencing Primers: Designing Primers Based on Sequences

3'Sequencing Primers: Designing Primers Based on Sequences

 

pBE2-R Description

The shuttle expression vector pBE2R was constructed by introducing strong promoter P43 from pWB980 plasmid and introducing signal peptide and propeptide of alkaline albumin enzyme from Bacillus alkalophilus downstream. The number of plasmid copies showed that pBE2R contained about one copy in each cell of E. coli and Bacillus subtilis. Plasmid stability test showed that the plasmid passed through 30 generations in E. coli and Bacillus subtilis without loss, and could be stable in E. coli and Bacillus subtilis.

 

pBE2-R Sequence

LOCUS       Exported                7014 bp ds-DNA     circular SYN 29-SEP-2017

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pBE2R

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 7014)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Sep 29, 2017  

FEATURES             Location/Qualifiers

     source          1..7014

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        803..907

                     /gene="bla"

                     /label=AmpR promoter

     CDS             908..1768

                     /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"

     rep_origin      1939..2527

                     /direction=RIGHT

                     /label=ori

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

                     replication"

     CDS             complement(3930..4700)

                     /codon_start=1

                     /label=KanR/NeoR

                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR

                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG

                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT

                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD

                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"

     CDS             complement(4860..5864)

                     /codon_start=1

                     /gene="repB"

                     /product="RepB replication protein"

                     /label=repB

                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"

                     /db_xref="GI:22652809"

                     /protein_id="AAN03827.1"

                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY

                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI

                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV

                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN

                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS

                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"

     misc_feature    6283..6587

                     /label=P43 promoter

     misc_feature    6620..6952

                     /label=Apr sp+pro

     CDS             6983..7000

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

ORIGIN

        1 tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc

       61 gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg aagcagaccc

      121 tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc ttctgagttt

      181 ctggtaacgc cgttccgcac cccggaaatg gtcagcgaac caatcagcag ggtcatcgct

      241 agccagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt

      301 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc

      361 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc

      421 gccatctcct tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg

      481 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg atatggtgca ctctcagtac

      541 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc

      601 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg

      661 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct

      721 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg

      781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc

      841 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag

      901 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg

      961 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt

     1021 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt

     1081 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt

     1141 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa

     1201 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag

     1261 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac

     1321 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac

     1381 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac

     1441 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac

     1501 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact

     1561 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg

     1621 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt

     1681 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat

     1741 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta

     1801 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa

     1861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga

     1921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac

     1981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt

     2041 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc

     2101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat

     2161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag

     2221 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc

     2281 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag

     2341 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac

     2401 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg

     2461 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct

     2521 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc

     2581 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga

     2641 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga

     2701 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg

     2761 cagctgattt cactttttgc attctacaaa ctgcataact catatgtaaa tcgctccttt

     2821 ttaggtggca caaatgtgag gcattttcgc tctttccggc aaccacttcc aagtaaagta

     2881 taacacacta tactttatat tcataaagtg tgtgctctgc gaggctgtcg gcagtgccga

     2941 ccaaaaccat aaaaccttta agacctttct tttttttacg agaaaaaaga aacaaaaaaa

     3001 cctgccctct gccacctcag caaagggggg ttttgctctc gtgctcgttt aaaaatcagc

     3061 aagggacagg tagtattttt tgagaagatc actcaaaaaa tctccacctt taaacccttg

     3121 ccaattttta ttttgtccgt tttgtctagc ttaccgaaag ccagactcag caagaataaa

     3181 atttttattg tctttcggtt ttctagtgta acggacaaaa ccactcaaaa taaaaaagat

     3241 acaagagagg tctctcgtat cttttattca gcaatcgcgc ccgattgctg aacagattaa

     3301 taatagattt tagcttttta tttgttgaaa aaagctaatc aaattgttgt cgggatcaat

     3361 tactgcaaag tctcgttcat cccaccactg atcttttaat gatgtattgg ggtgcaaaat

     3421 gcccaaaggc ttaatatgtt gatataattc atcaattccc tctacttcaa tgcggcaact

     3481 agcagtacca gcaataaacg actccgcacc tgtacaaacc ggtgaatcat tactacgaga

     3541 gcgccagctt catcacttgc ctcccataga tgaatccgaa cctcattaca cattagaact

     3601 gcgaatccat cttcatggtg aaccaaagtg aaacctagtt tatcgcaata aaaacctata

     3661 ctctttttaa tatccccgac tggcaatgcc gggatagact gtaacattct cacgcataaa

     3721 atcccctttc attttctaat gtaaatctat taccttatta ttaattcaat tcgctcataa

     3781 ttaatccttt ttcttattac gcaaaatggc ccgatttaag cacacccttt attccgttaa

     3841 tgcgccatga cagccatgat aattactaat actaggagaa gttaataaat acgtaaccaa

     3901 catgattaac aattattaga ggtcatcgtt caaaatggta tgcgttttga cacatccact

     3961 atatatccgt gtcgttctgt ccactcctga atcccattcc agaaattctc tagcgattcc

     4021 agaagtttct cagagtcgga aagttgacca gacattacga actggcacag atggtcataa

     4081 cctgaaggaa gatctgattg cttaactgct tcagttaaga ccgaagcgct cgtcgtataa

     4141 cagatgcgat gatgcagacc aatcaacatg gcacctgcca ttgctacctg tacagtcaag

     4201 gatggtagaa atgttgtcgg tccttgcaca cgaatattac gccatttgcc tgcatattca

     4261 aacagctctt ctacgataag ggcacaaatc gcatcgtgga acgtttgggc ttctaccgat

     4321 ttagcagttt gatacacttt ctctaagtat ccacctgaat cataaatcgg caaaatagag

     4381 aaaaattgac catgtgtaag cggccaatct gattccacct gagatgcata atctagtaga

     4441 atctcttcgc tatcaaaatt cacttccacc ttccactcac cggttgtcca ttcatggctg

     4501 aactctgctt cctctgttga catgacacac atcatctcaa tatccgaata gggcccatca

     4561 gtctgacgac caagagagcc ataaacacca atagccttaa catcatcccc atatttatcc

     4621 aatattcgtt ccttaatttc atgaacaatc ttcattcttt cttctctagt cattattatt

     4681 ggtccattca ctattctcat tcccttttca gataatttta gatttgcttt tctaaataag

     4741 aatatttgga gagcaccgtt cttattcagc tattaataac tcgtcttcct aagcatcctt

     4801 caatcctttt aataacaatt atagcatcta atcttcaaca aactggcccg tttgttgaac

     4861 tactctttaa taaaataatt tttccgttcc caattccaca ttgcaataat agaaaatcca

     4921 tcttcatcgg ctttttcgtc atcatctgta tgaatcaaat cgccttcttc tgtgtcatca

     4981 aggtttaatt ttttatgtat ttcttttaac aaaccaccat aggagattaa ccttttacgg

     5041 tgtaaacctt cctccaaatc agacaaacgt ttcaaattct tttcttcatc atcggtcata

     5101 aaatccgtat cctttacagg atattttgca gtttcgtcaa ttgccgattg tatatccgat

     5161 ttatatttat ttttcggtcg aatcatttga acttttacat ttggatcata gtctaatttc

     5221 attgcctttt tccaaaattg aatccattgt ttttgattca cgtagttttc tgtattctta

     5281 aaataagttg gttccacaca taccaataca tgcatgtgct gattataaga attatcttta

     5341 ttatttattg tcacttccgt tgcacgcata aaaccaacaa gatttttatt aattttttta

     5401 tattgcatca ttcggcgaaa tccttgagcc atatctgaca aactcttatt taattcttcg

     5461 ccatcataaa catttttaac tgttaatgtg agaaacaacc aacgaactgt tggcttttgt

     5521 ttaataactt cagcaacaac cttttgtgac tgaatgccat gtttcattgc tctcctccag

     5581 ttgcacattg gacaaagcct ggatttacaa aaccacactc gatacaactt tctttcgcct

     5641 gtttcacgat tttgtttata ctctaatatt tcagcacaat cttttactct ttcagccttt

     5701 ttaaattcaa gaatatgcag aagttcaaag taatcaacat tagcgatttt cttttctctc

     5761 catggtctca cttttccact ttttgtcttg tccactaaaa cccttgattt ttcatctgaa

     5821 taaatgctac tattaggaca cataatatta aaagaaaccc ccatctattt agttatttgt

     5881 ttagtcactt ataactttaa cagatggggt ttttctgtgc aaccaatttt aagggttttc

     5941 aatactttaa aacacataca taccaacact tcaacgcacc tttcagcaac taaaataaaa

     6001 atgacgttat ttctatatgt atcaagataa gaaagaacaa gttcaaaacc atcaaaaaaa

     6061 gacacctttt caggtgcttt ttttatttta taaactcatt ccctgatctc gacttcgttc

     6121 tttttttacc tctcggttat gagttagttc aaattcgttc tttttaggtt ctaaatcgtg

     6181 tttttcttgg aattgtgctg ttttatcctt taccttgtct acaaacccct taaaaacgtt

     6241 tttaaaggct tttaagccgt ctgtacgttc cttaaggaat tcgagctcag ctttattgag

     6301 tggatgatta tattcctttt gataggtggt atgttttcgc ttgaactttt aaatacagcc

     6361 attgaacata cggttgattt aataactgac aaacatcacc ctcttgctaa agcggccaag

     6421 gacgctgccg ccggggctgt ttgcgttttt gccgtgattt cgtgtattat tggtttactt

     6481 atttttttgc caaagctgta atggctgaaa attcttacat ttattttaca tttttagaaa

     6541 tgggcgtgaa aaaaagcgcg cgattatgta aaatataaag tgatagcggt accattatag

     6601 gtaagagagg aatgtacaca tgaagaaacc gttggggaaa attgtcgcaa gcaccgcact

     6661 actcatttct gttgctttta gttcatcgat cgcatcggct gctgaagaag caaaagaaaa

     6721 atatttaatt ggctttaatg agcaggaagc tgtcagtgag tttgtagaac aagtagaggc

     6781 aaatgacgag gtcgccattc tctctgagga agaggaagtc gaaattgaat tgcttcatga

     6841 atttgaaacg attcctgttt tatccgttga gttaagccca gaagatgtgg acgcgcttga

     6901 actcgatcca gcgatttctt atattgaaga ggatgcagaa gtaacgacaa tgggatcctc

     6961 tagagtcgac ctgcagctcg agcaccacca ccaccaccac aagcttgtat tcta

//


Caution:
Product is for research use only!
 

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