LOCUS pMAL-p4X 6720 bp DNA circular 06-JUN-2014
DEFINITION Cloning vector pMAL-p4X, complete sequence.
ACCESSION
VERSION
KEYWORDS .
SOURCE Cloning vector pMAL-p4X
ORGANISM Cloning vector pMAL-p4X
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 6720)
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) In
press
REFERENCE 2 (bases 1 to 6720)
AUTHORS Riggs,P.D.
TITLE Direct Submission
JOURNAL Submitted (22-OCT-2007) Research Department, New England Biolabs,
240 County Road, Ipswich, MA 01938, USA
FEATURES Location/Qualifiers
source 1..6720
/organism="Cloning vector pMAL-p4X"
/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..2991
/gene="malE expression cassette"
/note="malE sequence 1528..2703; multiple cloning site
region 2704..2809; lacZalpha sequence 2810..2991"
/codon_start=1
/product="maltose binding protein (MBP) fusion"
/translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYN
GLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLL
AEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPA
LDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGL
TFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTF
KGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYE
EELVKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT
NSSSNNNNNNNNNNLGIEGRISEFGSSRVDLQASLALAVVLQRRDWENPGVTQLNRLA
AHPPFASWRNSEEARTDRPSQQLRSLNGEWQLGCFGG"
primer_bind 2662..2685
/note="malE primer sequence"
misc_feature 2770..2807
/gene="malE expression cassette"
/note="multiple cloning site (EcoRI-HindIII)"
terminator 3179..3382
/note="rrnB T1T2 transcription terminators"
gene 3492..4352
/gene="bla"
CDS 3492..4352
/gene="bla"
/note="ampR (confers resistance to ampicillin)"
/codon_start=1
/product="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
sig_peptide 3492..3560
/gene="bla"
/note="required for secretion to the periplasm; cleaved
off to form the mature beta-lactamase protein"
rep_origin complement(4394..4907)
/note="M13 origin of replication (+ -)"
rep_origin 5018..5606
/note="pMB1 origin of replication (clockwise) (RNAII -35
to RNA/DNA switch point)"
gene complement(6036..6227)
/gene="rop"
CDS complement(6036..6227)
/gene="rop"
/exception="alternative start codon"
/codon_start=1
/transl_table=11
/product="ROP protein"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH
ADELYRSCLARFGDDGENL"
BASE COUNT 1714 a 1692 c 1783 g 1531 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 aagagttggt gaaagatccg cggattgccg ccactatgga aaacgcccag
2581 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
2641 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag
2701 actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg gatcgaggga
2761 aggatttcag aattcggatc ctctagagtc gacctgcagg caagcttggc actggccgtc
2821 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
2881 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
2941 cagttgcgca gcctgaatgg cgaatggcag cttggctgtt ttggcggatg agataagatt
3001 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
3061 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
3121 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
3181 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
3241 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
3301 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
3361 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
3421 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
3481 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
3541 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
3601 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
3661 gttttcgccc cgaagaacgt ttcccaatga tgagcacttt taaagttctg ctatgtggcg
3721 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
3781 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
3841 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
3901 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
3961 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
4021 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
4081 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
4141 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
4201 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
4261 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
4321 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
4381 tttagattga tttaccccgg ttgataatca gaaaagcccc aaaaacagga agattgtata
4441 agcaaatatt taaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
4501 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
4561 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
4621 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
4681 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
4741 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
4801 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
4861 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtaaa aggatctagg
4921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
4981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
5041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
5101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
5161 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
5221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
5281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
5341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
5401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
5461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
5521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
5581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
5641 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
5701 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
5761 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
5821 tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc
5881 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
5941 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
6001 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
6061 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg
6121 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
6181 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
6241 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
6301 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
6361 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
6421 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
6481 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
6541 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
6601 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
6661 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgaaatt
//