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pMBPC载体基本信息
pMBPC载体图谱质粒图谱和多克隆位点信息
pMBPC载体简介
pMBP-C 表达载体采用 E.coli tac 启动子和胞内可溶性麦芽糖结合蛋白（maltose binding protein），同时选用凝血酶切割位点并在 C端添加6 个His便于采用 chelating sepharose 亲和层析纯化目标蛋白。该表达系统在 Medline 里有大量背景文献参考。它通过 MBP胞内融合表达，表达产物可占菌体的 20%-30%，可形成可溶性蛋白，也可形成包涵体。融合蛋白还可以通过 Amylose 亲和层析纯化。
pMBPC载体序列
LOCUS pMBP-C 6666 bp ds-DNA circular SYN 05-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMBP-C
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6666)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6666
/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..2628
/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
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
CDS 2728..2745
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
primer_bind complement(2756..2772)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
terminator 3121..3207
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3299..3326
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3346..3437
/gene="bla"
/note="AmpR promoter"
CDS 3438..4298
/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(4340..4853)
/direction=LEFT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 4964..5552
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 5738..5880
/note="bom"
/note="basis of mobility region from pBR322"
CDS complement(5982..6173)
/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 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 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg
2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctct
2641 aacaacaaca acaacaacaa caataacaac ctgggtatcg atactactga gaactatccg
2701 cgtggatccg cagaattcag cgctagccac catcatcatc atcattaagc ttggcactgg
2761 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
2821 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
2881 cccaacagtt gcgcagcctg aatggcgaat ggcagcttgg ctgttttggc ggatgagata
2941 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
3001 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
3061 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
3121 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
3181 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
3241 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
3301 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctctttttgt ttatttttct
3361 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
3421 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
3481 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
3541 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
3601 ttgagagttt tcgccccgaa gaacgttctc caatgatgag cacttttaaa gttctgctat
3661 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
3721 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
3781 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
3841 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
3901 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
3961 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
4021 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
4081 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
4141 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
4201 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
4261 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
4321 atatacttta gattgattta ccccggttga taatcagaaa agccccaaaa acaggaagat
4381 tgtataagca aatatttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
4441 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
4501 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
4561 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
4621 acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
4681 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
4741 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
4801 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga
4861 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
4921 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
4981 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
5041 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
5101 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
5161 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
5221 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
5281 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
5341 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
5401 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
5461 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
5521 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
5581 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
5641 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
5701 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
5761 cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga
5821 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
5881 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
5941 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
6001 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg cagcgattca
6061 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
6121 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
6181 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
6241 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
6301 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
6361 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
6421 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
6481 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
6541 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
6601 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
6661 gaaatt
//
