pMal-T-Avi-His/BirA

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  • Model: PVT12239
  • 20 Units in Stock
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pMal-T-Avi-His/BirA

PVT12239    2ug
 

 

pMal-T-Avi-His/BirA Search name

pMal-T-Avi-His/BirA,pMal-T-Avi-His/BirA vector,pMal-T-Avi-His/BirA plasmid,pMal-T-Avi-His/BirA map

 

 

pMal-T-Avi-His/BirA Information

Promoter: TAC

Prokaryotic resistance: amp

Clone strain: DH5 α

Culture condition: 37 ℃

Plasmid host: Escherichia coli

Purpose of plasmid: protein expression

Fragment type: gene CDs

Fragment species: empty body

Prokaryotic resistance: ampicillin

pMal-T-Avi-His-BirA-vector

 

pMal-T-Avi-His/BirA Sequence

LOCUS       Exported                7292 bp ds-DNA     circular SYN 10-JUL-2018

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pMal-T-Avi-His-BirA

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 7292)

  AUTHORS   .

  TITLE     Direct Submission

FEATURES             Location/Qualifiers

     source          1..7292

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     CDS             complement(101..292)

                     /codon_start=1

                     /gene="rop"

                     /product="Rop protein, which maintains plasmids at low copy

                     number"

                     /note="rop"

                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA

                     DELYRSCLARFGDDGENL"

     promoter        788..865

                     /gene="lacI (mutant)"

                     /note="lacIq promoter"

                     /note="In the lacIq allele, a single base change in the 

                     promoter boosts expression of the lacI gene about 10-fold."

     CDS             866..1948

                     /codon_start=1

                     /gene="lacI"

                     /product="lac repressor"

                     /note="lacI"

                     /note="The lac repressor binds to the lac operator to 

                     inhibit transcription in E. coli. This inhibition can be 

                     relieved by adding lactose or 

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL

                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV

                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH

                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA

                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC

                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR

                     ALADSLMQLARQVSRLESGQ"

     promoter        2191..2219

                     /note="tac promoter"

                     /note="strong E. coli promoter; hybrid between the trp and 

                     lac UV5 promoters"

     protein_bind    2227..2243

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="lac operator"

                     /note="The lac repressor binds to the lac operator to 

                     inhibit transcription in E. coli. This inhibition can be 

                     relieved by adding lactose or 

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     CDS             2313..3413

                     /codon_start=1

                     /gene="malE (mutated)"

                     /product="maltose binding protein from E. coli"

                     /note="MBP"

                     /note="This version of the gene does not encode a signal 

                     sequence, so MBP will remain in the cytosol."

                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE

                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL

                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA

                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE

                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL

                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF

                     WYAVRTAVINAASGRQTVDEALKDAQT"

     CDS             3462..3473

                     /codon_start=1

                     /product="Factor Xa recognition and cleavage site"

                     /note="Factor Xa site"

                     /translation="IEGR"

     CDS             3551..3571

                     /codon_start=1

                     /product="tobacco etch virus (TEV) protease recognition and

                     cleavage site"

                     /note="TEV site"

                     /translation="EDLYFQS"

     misc_feature    3590..3631

                     /note="AviTag"

     CDS             3638..3655

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /note="6xHis"

                     /translation="HHHHHH"

     CDS             3664..3687

                     /codon_start=1

                     /product="FLAG(R) epitope tag, followed by an enterokinase 

                     cleavage site"

                     /note="FLAG"

                     /translation="DYKDDDDK"

     terminator      3699..3785

                     /gene="Escherichia coli rrnB"

                     /note="T1"

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

                     gene"

     terminator      3877..3904

                     /note="T2"

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

                     gene"

     promoter        3932..4022

                     /gene="bla"

                     /note="Amp promoter"

     CDS             4023..4883

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /note="Amp"

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      4971..5559

                     /direction=RIGHT

                     /note="ori"

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

                     replication"

     protein_bind    5847..5871

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="lac operator"

                     /note="The lac repressor binds to the lac operator to 

                     inhibit transcription in E. coli. This inhibition can be 

                     relieved by adding lactose or 

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     RBS             5886..5908

                     /note="efficient ribosome binding site from bacteriophage 

                     T7 gene 10 (Olins and Rangwala, 1989)"

     CDS             5915..6877

                     /codon_start=1

                     /gene="birA"

                     /product="E. coli biotin protein ligase"

                     /note="BirA"

                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD

                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG

                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK

                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG

                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG

                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"

     terminator      7000..7047

                     /note="T7 terminator"

                     /note="transcription terminator for bacteriophage T7 RNA 

                     polymerase"

ORIGIN

        1 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg

       61 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat

      121 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac

      181 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct

      241 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct

      301 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc

      361 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac

      421 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct

      481 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc

      541 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac

      601 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg

      661 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc

      721 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg

      781 aaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga

      841 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta

      901 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc

      961 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt

     1021 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc

     1081 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc

     1141 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca

     1201 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga

     1261 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga

     1321 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga

     1381 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt

     1441 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc

     1501 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat

     1561 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg

     1621 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatattt cggtagtggg

     1681 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga

     1741 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc

     1801 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc

     1861 caatacacaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca

     1921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc

     1981 attaggcaca attctcatgt ttgacagctt atcatcgact gcacggtgca ccaatgcttc

     2041 tggcgtcagg cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata

     2101 attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa

     2161 cggttctggc aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt

     2221 gtggaattgt gagcggataa caatttcaca caggaaacag ccagtccgtt taggtgtttt

     2281 cacgagcaat tgaccaacaa ggaccataga ttatgaaaat cgaagaaggt aaactggtaa

     2341 tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag aaattcgaga

     2401 aagataccgg aattaaagtc accgttgagc atccggataa actggaagag aaattcccac

     2461 aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac cgctttggtg

     2521 gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc caggacaagc

     2581 tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct tacccgatcg

     2641 ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg ccaaaaacct

     2701 gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt

     2761 tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg ggttatgcgt

     2821 tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac gctggcgcga

     2881 aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat gcagacaccg

     2941 attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc atcaacggcc

     3001 cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg gtactgccga

     3061 ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt attaatgccg

     3121 ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg actgatgaag

     3181 gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg

     3241 aagagttggt gaaagatccg cgtattgccg ccactatgga aaacgcccag aaaggtgaaa

     3301 tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact gcggtgatca

     3361 acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag actaattcga

     3421 gttctaacaa caacaacaat aacaataaca acaacctcgg gatcgaggga aggatttcac

     3481 atatgtccat gggcggccgc gatatcgtcg acaagaggtg atatcgatcg gagctctcga

     3541 gccaaccact gaggatctgt actttcagag cgataacgat ggatccctag gactgaacga

     3601 catcttcgaa gctcagaaaa tcgaatggca cggatcccat catcaccatc accactagaa

     3661 ttcgattaca aggatgacga cgataagtaa ataagcttca aataaaacga aaggctcagt

     3721 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga

     3781 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag

     3841 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc

     3901 tttttgcgtt tctacaaact ctttcggtcc gttgtttatt tttctaaata cattcaaata

     3961 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga

     4021 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc

     4081 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg

     4141 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc

     4201 ccgaagaacg tttcccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat

     4261 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact

     4321 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat

     4381 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga

     4441 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc

     4501 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga

     4561 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag

     4621 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc

     4681 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt

     4741 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct

     4801 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg

     4861 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg

     4921 atttccttag gactgagcgt caaccccgta gaaaagatca aaggatcttc ttgagatcct

     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt

     5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg

     5101 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct

     5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc

     5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg

     5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa

     5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg

     5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg

     5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga

     5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt

     5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tccggtaccg cgcccaacag

     5641 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa

     5701 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc

     5761 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc

     5821 gcgaaattaa tacgactcac tagaggggaa ttgtgagcgg ataacaattc ccctctagaa

     5881 ataattttgt ttaactttaa gaaggagata taccatgaag gataacaccg tgccactgaa

     5941 attgattgcc ctgttagcga acggtgaatt tcactctggc gagcagttgg gtgaaacgct

     6001 gggaatgagc cgggcggcta ttaataaaca cattcagaca ctgcgtgact ggggcgttga

     6061 tgtctttacc gttccgggta aaggatacag cctgcctgag cctatccagt tacttaatgc

     6121 taaacagata ttgggtcagc tggatggcgg tagtgtagcc gtgctgccag tgattgactc

     6181 cacgaatcag taccttcttg atcgtatcgg agagcttaaa tcgggcgatg cttgcattgc

     6241 agaataccag caggctggcc gtggtcgccg gggtcggaaa tggttttcgc cttttggcgc

     6301 aaacttatat ttgtcgatgt tctggcgtct ggaacaaggc ccggcggcgg cgattggttt

     6361 aagtctggtt atcggtatcg tgatggcgga agtattacgc aagctgggtg cagataaagt

     6421 tcgtgttaaa tggcctaatg acctctatct gcaggatcgc aagctggcag gcattctggt

     6481 ggagctgact ggcaaaactg gcgatgcggc gcaaatagtc attggagccg ggatcaacat

     6541 ggcaatgcgc cgtgttgaag agagtgtcgt taatcagggg tggatcacgc tgcaggaagc

     6601 ggggatcaat ctcgatcgta atacgttggc ggccatgcta atacgtgaat tacgtgctgc

     6661 gttggaactc ttcgaacaag aaggattggc accttatctg tcgcgctggg aaaagctgga

     6721 taattttatt aatcgcccag tgaaacttat cattggtgat aaagaaatat ttggcatttc

     6781 acgcggaata gacaaacagg gggctttatt acttgagcag gatggaataa taaaaccctg

     6841 gatgggcggt gaaatatccc tgcgtagtgc agaaaaaggt aagctttgat gaggactagt

     6901 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga

     6961 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc

     7021 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atgctagcac

     7081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga

     7141 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg

     7201 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata

     7261 taaggtgcac tgtgactggg tcatggctgc gc

//
 

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