pMal-p2E
PVT0404 2ug Lead time 3-4 weeks
pMal-p2E Information
Promoter: Tac
Replicon: ColE1 ori
Terminator: rrnB T1 Terminator
Plasmid classification: Escherichia coli vector; pMal series expression plasmid
Plasmid size: 6724bp
Plasmid label: N-MBP, N-EK
Prokaryotic resistance: ampicillin Amp
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: BL21 (DE3)
Culture conditions: 37 centigrade, aerobic, LB
Inducement: IPTG or lactose and its analogues
5'primers: MalE primers: 5-GGTCGTCAGACTGTCGATGAAGCC-3; MBP-F: 5-gatgaagccctgaaagacgcgcag-3
3'sequencing primers: pBad-R: 5-gatttaatctgtatcagg-3; M13-F: 5-TGTAAAACGACGGCCAGT-3
Remarks: fusion expression of maltose binding protein MBP, protein located in periplasmic
pMal-p2E Description
PMAL is an efficient protein fusion expression and purification system. The pMAL vector contains the malE gene of the Escherichia coli that encodes the Maltose Binding Protein (MBP). The downstream polyclonal loci are convenient for the insertion of the target gene and express the fusion protein with MBP on the N end. The cloning gene was highly expressed through the "TAC" strong promoter and the malE translation initiation signal, and the affinity purification of maltose by MBP was further used to achieve one step affinity purification of the fusion protein by Amylose column.
pMal-p2E Sequence
LOCUS Exported 6724 bp ds-DNA circular SYN 12-08-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMal-p2E
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6724)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-12 from SnapGene Viewer 2.8.1
FEATURES Location/Qualifiers
source 1..6724
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 3..80
/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 81..1163
/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"
misc_feature 1406..1434
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1442..1458
/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 1528..2703
/codon_start=1
/gene="malE"
/product="maltose binding protein from E. coli"
/note="MBP"
/note="This gene encodes a signal sequence that can direct
MBP to the bacterial periplasm."
/translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKD
PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
CDS 2752..2766
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
primer_bind complement(2814..2830)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
terminator 3179..3265
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3357..3384
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3404..3495
/gene="bla"
/note="AmpR promoter"
CDS 3496..4356
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(4398..4911)
/direction=LEFT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 5022..5610
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 5796..5938
/note="bom"
/note="basis of mobility region from pBR322"
CDS complement(6040..6231)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
1501 gcacttcacc aacaaggacc atagcatatg aaaataaaaa caggtgcacg catcctcgca
1561 ttatccgcat taacgacgat gatgttttcc gcctcggctc tcgccaaaat cgaagaaggt
1621 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag
1681 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag
1741 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac
1801 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc
1861 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct
1921 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg
1981 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc
2041 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg
2101 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac
2161 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat
2221 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc
2281 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg
2341 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt
2401 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg
2461 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag
2521 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccactatgga aaacgcccag
2581 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
2641 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag
2701 actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg ggatgacgat
2761 gacaaggtac cggaattcgg atcctctaga gtcgacctgc aggcaagctt ggcactggcc
2821 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
2881 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
2941 caacagttgc gcagcctgaa tggcgaatgg cagcttggct gttttggcgg atgagataag
3001 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
3061 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
3121 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
3181 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
3241 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
3301 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
3361 ggccatcctg acggatggcc tttttgcgtt tctacaaact ctttttgttt atttttctaa
3421 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
3481 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
3541 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
3601 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
3661 gagagttttc gccccgaaga acgttctcca atgatgagca cttttaaagt tctgctatgt
3721 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
3781 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
3841 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
3901 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
3961 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
4021 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
4081 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
4141 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
4201 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
4261 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
4321 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
4381 atactttaga ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg
4441 tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
4501 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
4561 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
4621 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
4681 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
4741 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
4801 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
4861 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc
4921 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
4981 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
5041 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
5101 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
5161 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
5221 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
5281 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
5341 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
5401 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
5461 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
5521 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
5581 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
5641 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
5701 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
5761 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
5821 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
5881 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
5941 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
6001 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
6061 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca
6121 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
6181 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
6241 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
6301 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
6361 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
6421 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
6481 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
6541 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
6601 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
6661 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
6721 aatt
//
Caution:
Product is for research use only!
Search name
pMal-p2E,Plasmid pMal-p2E,pMal-p2E vector