| 出品公司: | -- |
|---|---|
| 载体名称: | pCold-MBP |
| 质粒类型: | 大肠杆菌载体 |
| 高拷贝/低拷贝: | -- |
| 克隆方法: | -- |
| 启动子: | CspA |
| 载体大小: | 5505 bp |
| 5' 测序引物及序列: | pCold-F: 5-ACGCCATATCGCCGAAAGG-3 |
| 3' 测序引物及序列: | pCold-R: 5-GGCAGGGATCTTAGATTCTG-3 |
| 载体标签: | N-Factor Xa，6*His |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | -- |
| 克隆菌株: | DH5α 等 |
| 宿主细胞（系）: | -- |
| 备注: | -- |
| 产品目录号: | -- |
| 稳定性: | -- |
| 组成型/诱导型: | -- |
| 病毒/非病毒: | -- |
买家导航
pColdMBP载体基本信息
pColdMBP载体图谱质粒图谱和多克隆位点信息
pColdMBP载体简介
pColdMBP载体序列
LOCUS pCold MBP 5505 bp ds-DNA circular SYN 22-七月-2016
DEFINITION synthetic circular DNA
KEYWORDS pCold MBP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5505)
TITLE Direct Submission
http://www.biofeng.com/
FEATURES Location/Qualifiers
source 1..5505
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 15..81
/note="cspA promoter"
protein_bind 84..100
/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)."
misc_feature 122..255
/note="cspA 5'UTR"
misc_feature 256..270
/note="TEE"
misc_feature 256..258
/note="M"
CDS 271..288
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 289..300
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
CDS 304..1404
/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
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
misc_feature 1405..1458
/note="multiple cloning site"
misc_feature 1466..1610
/note="cspA 3'UTR"
misc_feature 1494..1511
/note="transcription terminator"
rep_origin complement(1809..2264)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2364..2468
/gene="bla"
/note="AmpR promoter"
CDS 2469..3329
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3500..4088
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4260..5342)
/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"
promoter complement(5343..5420)
/gene="lacI"
/note="lacI promoter"
ORIGIN
1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa
61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat
121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt
181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt
241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcatat cgaaggtagg
301 catatgaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
361 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
421 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
481 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
541 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
601 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
661 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
721 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
781 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
841 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
901 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
961 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
1021 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
1081 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
1141 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
1201 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc
1261 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
1321 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
1381 gaagccctga aagacgcgca gactgagctc ggtaccctcg agggatccga attcaagctt
1441 gtcgacctgc agtctagata ggtaatctct gcttaaaagc acagaatcta agatccctgc
1501 catttggcgg ggattttttt atttgttttc aggaaataaa taatcgatcg cgtaataaaa
1561 tctattatta tttttgtgaa gaataaattt gggtgcaatg agaatgcgca ggccctttcg
1621 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
1681 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
1741 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
1801 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
1861 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
1921 gcccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
1981 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
2041 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
2101 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
2161 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
2221 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
2281 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgtcag
2341 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
2401 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
2461 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
2521 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
2581 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
2641 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
2701 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
2761 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
2821 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
2881 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
2941 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
3001 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
3061 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
3121 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
3181 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
3241 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
3301 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
3361 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
3421 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
3481 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
3541 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
3601 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
3661 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
3721 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
3781 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
3841 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
3901 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
3961 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
4021 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
4081 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
4141 ctcacatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
4201 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
4261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
4321 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
4381 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
4441 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
4501 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
4561 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
4621 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
4681 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
4741 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
4801 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
4861 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
4921 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
4981 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
5041 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
5101 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
5161 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
5221 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
5281 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
5341 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
5401 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
5461 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgc
//
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