| 质粒名称: | pET MBP His6 LIC cloning vector (2Cc-T) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 37237 |
| 存储实验室: | Scott Gradia |
|
载体骨架基本信息 |
|
| 载体名称: | pET |
| 载体出品公司: | Clontech |
| 空载体大小: | 5898 bp |
| 完整质粒大小: | 4989 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 启动子: | T7 |
| 标签/融合蛋白: |
MBP (C terminal on backbone)
His6 (C terminal on backbone)
|
| 克隆方法: | Ligation Independent Cloning |
| 5’克隆位点: | T7 promoter |
| 3’克隆位点: | MBP 226R |
| 5’测序引物 | ttgagcgtagccaccaaag |
|
文献引用方法 |
|
| 材料和方法部分: | pET MBP His6 LIC cloning vector (2Cc-T) was a gift from Scott Gradia (Addgene plasmid # 37237 ; http://n2t.net/addgene:37237 ; RRID:Addgene_37237) |
| 参考文献部分: | |
买家导航
pET MBP His6 LIC cloning vector (2Cc-T)载体质粒基本信息
pET MBP His6 LIC cloning vector (2Cc-T)质粒图谱载体图谱和pET MBP His6 LIC cloning vector (2Cc-T)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pET MBP His6 LIC cloning vector (2Cc-T)质粒载体简介
pET MBP His6 LIC cloning vector (2Cc-T)质粒序列载体序列
LOCUS pET MBP His6 LIC cloning vector (2Cc-T) 5895 bp DNA circular SYN 14-APR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5895)
TITLE 2-series C-terminal fusion vectors for E. coli expression
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5895)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT Sequence Label: pET MBP His6 LIC cloning vector (2Cc-T)
FEATURES Location/Qualifiers
source 1..5895
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(18..36)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 104..208
/gene="bla"
/label=AmpR promoter
CDS join(209..277,278..1069)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(427..446)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1240..1828
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1729..1748
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1982..1999
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2014..2156
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2082..2101)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2242..2264
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2258..2449)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 3907..3926
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 4010..4029
/label=T7
/note="T7 promoter, forward primer"
promoter 4010..4028
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 4117..4139
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 4180..5280
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=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"
primer_bind 5254..5277
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 5290..5307
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5431..5449)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5445..5492
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(join(5858..5863,5864..5880,5881..5886))
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
ORIGIN
1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc
3721 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
3781 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
3841 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
3901 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
3961 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
4021 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa
4081 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat
4141 atagttaacc tctacttcca atccggctct agcggctcta tgaaaatcga agaaggtaaa
4201 ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa
4261 ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa
4321 ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc
4381 tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag
4441 gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac
4501 ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca
4561 aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg
4621 ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt
4681 tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct
4741 ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca
4801 gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc
4861 aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta
4921 ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt
4981 aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact
5041 gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct
5101 tacgaggaag agttggcgaa agatccacgt attgccgcca ctatggaaaa cgcccagaaa
5161 ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg
5221 gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact
5281 ggcagtagcc accatcacca tcaccattaa taaccaactc cataaggatc cgcgatcgcg
5341 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
5401 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
5461 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggacatc
5521 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
5581 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
5641 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
5701 tggacgacat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
5761 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
5821 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
5881 tgtcaaacat gagaa
//
