LOCUS pMAL-p2E 6723 bp DNA circular 06-JUN-2014
DEFINITION Cloning vector pMAL-p2E, complete sequence.
ACCESSION
VERSION
KEYWORDS .
SOURCE Cloning vector pMAL-p2E
ORGANISM Cloning vector pMAL-p2E
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 6723)
AUTHORS Riggs,P.
TITLE Expression and purification of maltose-binding protein fusions
JOURNAL (in) Asubel,F.A., Brent,R., Kingston,R.E., Moore,D.D.,
Seidman,J.G., Smith,J.A. and Struhl,K. (Eds.);
CURRENT PROTOCOLS IN MOLECULAR BIOLOGY: 1-14;
Greene Publishing and Wiley Interscience, New York, NY (1992)
REFERENCE 2 (bases 1 to 6723)
AUTHORS Riggs,P.D.
TITLE Direct Submission
JOURNAL Submitted (07-DEC-2007) Research Department, New England Biolabs,
240 County Road, Ipswich, MA 01938, USA
FEATURES Location/Qualifiers
source 1..6723
/organism="Cloning vector pMAL-p2E"
/mol_type="other DNA"
gene 15..1163
/gene="lacIq"
promoter 15..46
/gene="lacIq"
/note="lacIq promoter (overexpressing mutant) (clockwise)"
CDS 81..1163
/gene="lacIq"
/exception="alternative start codon"
/codon_start=1
/transl_table=11
/product="lac repressor (LacI)"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
QTASPRALADSLMQLARQVSRLESGQ"
gene 1406..3307
/gene="malE expression cassette"
promoter 1406..1433
/gene="malE expression cassette"
/note="Ptac promoter (clockwise)"
CDS 1528..2994
/gene="malE expression cassette"
/note="malE sequence 1528..2703; multiple cloning site
region 2704..2812; lacZalpha sequence 2813..2994"
/codon_start=1
/product="maltose binding protein (MBP) fusion"
/translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYN
GLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLL
AEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPA
LDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGL
TFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTF
KGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYE
EELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT
NSSSNNNNNNNNNNLGDDDDKVPEFGSSRVDLQASLALAVVLQRRDWENPGVTQLNRL
AAHPPFASWRNSEEARTDRPSQQLRSLNGEWQLGCFGG"
primer_bind 2662..2685
/note="malE primer sequence"
misc_feature 2773..2810
/gene="malE expression cassette"
/note="multiple cloning site (EcoRI-HindIII)"
terminator 3182..3385
/note="rrnB T1T2 transcription terminators"
gene 3495..4355
/gene="bla"
CDS 3495..4355
/gene="bla"
/note="ampR (confers resistance to ampicillin)"
/codon_start=1
/product="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
sig_peptide 3495..3563
/gene="bla"
/note="required for secretion to the periplasm; cleaved
off to form the mature beta-lactamase protein"
rep_origin complement(4397..4910)
/note="M13 origin of replication (+ -)"
rep_origin 5021..5609
/note="pMB1 origin of replication (clockwise) (RNAII -35
to RNA/DNA switch point)"
gene complement(6039..6230)
/gene="rop"
CDS complement(6039..6230)
/gene="rop"
/exception="alternative start codon"
/codon_start=1
/transl_table=11
/product="ROP protein"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
ADELYRSCLARFGDDGENL"
BASE COUNT 1716 a 1695 c 1782 g 1530 t
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 cttttgttta tttttctaaa
3421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
3481 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
3541 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
3601 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
3661 agagttttcg ccccgaagaa cgtttcccaa tgatgagcac ttttaaagtt ctgctatgtg
3721 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
3781 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
3841 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
3901 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
3961 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
4021 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
4081 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
4141 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
4201 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
4261 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
4321 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
4381 tactttagat tgatttaccc cggttgataa tcagaaaagc cccaaaaaca ggaagattgt
4441 ataagcaaat atttaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
4501 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
4561 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
4621 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
4681 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
4741 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
4801 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
4861 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt aaaaggatct
4921 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
4981 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
5041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
5101 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
5161 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
5221 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
5281 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
5341 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
5401 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
5461 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
5521 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
5581 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
5641 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
5701 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
5761 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
5821 atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc
5881 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
5941 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
6001 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
6061 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag
6121 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
6181 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
6241 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
6301 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
6361 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
6421 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
6481 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
6541 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
6601 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
6661 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgaa
6721 att
//