LOCUS pMAL-c5E 5680 bp DNA circular 12-MAR-2014
DEFINITION Cloning vector pMAL-c5E, complete sequence.
ACCESSION
VERSION
KEYWORDS .
SOURCE Cloning vector pMAL-c5E
ORGANISM Cloning vector pMAL-c5E
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 5680)
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 5680)
AUTHORS Riggs,P.D.
TITLE Direct Submission
JOURNAL Submitted (13-NOV-2008) Research Department, New England Biolabs,
240 County Road, Ipswich, MA 01938, USA
FEATURES Location/Qualifiers
source 1..5680
/organism="Cloning vector pMAL-c5E"
/mol_type="other DNA"
gene 15..1163
/gene="lacIq"
-35_signal 15..19
/gene="lacIq"
/note="lacIq promoter (clockwise); TGGTG"
-10_signal 40..46
/gene="lacIq"
/note="lacIq promoter (clockwise); CATGATA"
CDS 81..1163
/gene="lacIq"
/codon_start=1
/transl_table=11
/product="lac repressor (LacI)"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
QTASPRALADSLMQLARQVSRLESGQ"
gene 1406..2971
/gene="malE expression cassette"
-35_signal 1406..1411
/gene="malE expression cassette"
/note="Ptac promoter (clockwise); TTGACA"
-10_signal 1428..1433
/gene="malE expression cassette"
/note="Ptac promoter (clockwise); TATAAT"
CDS 1528..2757
/gene="malE expression cassette"
/note="malE sequence 1528..2628; spacer and enterokinase
site 2629..2690; multiple cloning site region 2691..2757"
/codon_start=1
/product="maltose binding protein (MBP) fusion"
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL
EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG
KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP
LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF
NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL
AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNI
PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGDDDDKVPHM
SMGGRDIVDGSEFPAGN"
primer_bind 2587..2610
/note="malE primer sequence"
misc_feature 2691..2750
/gene="malE expression cassette"
/note="multiple cloning site (KpnI-SbfI)"
terminator 2769..2812
/gene="malE expression cassette"
/note="rrnB T1 transcription terminator"
primer_bind complement(2826..2849)
/note="MCS reverse primer sequence"
terminator 2944..2971
/gene="malE expression cassette"
/note="rrnB T2 transcription terminator"
gene 3020..3950
/gene="bla"
-35_signal 3020..3025
/gene="bla"
/note="bla promoter (clockwise); TTCAAA"
-10_signal 3043..3048
/gene="bla"
/note="bla promoter (clockwise); GACAAT"
CDS 3090..3950
/gene="bla"
/note="ampR (confers resistance to ampicillin)"
/codon_start=1
/product="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
rep_origin 4038..4626
/note="pMB1 origin of replication (clockwise) (RNAII -35
to RNA/DNA switch point)"
gene complement(4996..5187)
/gene="rop"
CDS complement(4996..5187)
/gene="rop"
/codon_start=1
/transl_table=11
/product="ROP protein"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
ADELYRSCLARFGDDGENL"
BASE COUNT 1419 a 1432 c 1524 g 1305 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 tatttcggta 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 gcaattgacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg
1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
2461 ttggtgaaag atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg
2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
2641 aacaacaaca acaataacaa taacaacaac ctcggggatg acgatgacaa ggtaccgcat
2701 atgtccatgg gcggccgcga tatcgtcgac ggatccgaat tccctgcagg taattaaata
2761 agcttcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
2821 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
2881 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
2941 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt tcggtccgtt
3001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
3061 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
3121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
3181 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
3241 gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta
3301 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
3361 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
3421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
3481 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
3541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
3601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
3661 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
3721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
3781 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
3841 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
3901 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
3961 aagtttactc atatatactt tagattgatt tccttaggac tgagcgtcaa ccccgtagaa
4021 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
4081 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
4141 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
4201 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
4261 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
4321 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
4381 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
4441 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
4501 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
4561 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
4621 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
4681 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
4741 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
4801 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
4861 atataaggtg cactgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
4921 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
4981 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
5041 gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc
5101 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
5161 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
5221 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
5281 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
5341 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
5401 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
5461 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
5521 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
5581 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
5641 catgcgcacc cgtggccagg acccaacgct gcccgaaatt
//