| 质粒名称: | pET15-MHL |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 26092 |
| 存储实验室: | Cheryl Arrowsmith |
|
载体骨架基本信息 |
|
| 载体名称: | pET15-MHL |
| 载体出品公司: | SGC |
| 空载体大小: | 7737bp |
| 完整质粒大小: | 7734bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 启动子: | T7-lacO (lactose/IPTG inducible) |
| 标签/融合蛋白: |
6xHis (C terminal on backbone)
TEV cleavage site (C terminal on backbone)
|
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | T7 |
| 3’测序引物 | T7-term |
|
文献引用方法 |
|
| 材料和方法部分: | pET15-MHL was a gift from Cheryl Arrowsmith (Addgene plasmid # 26092 ; http://n2t.net/addgene:26092 ; RRID:Addgene_26092) |
买家导航
pET15-MHL载体质粒基本信息
pET15-MHL质粒图谱载体图谱和 pET15-MHL载体序列质粒序列多克隆位点信息
pET15-MHL质粒载体简介
pET15-MHL质粒序列载体序列
LOCUS pET15-MHL 7734 bp DNA circular SYN 14-APR-2025
DEFINITION SGC Empty backbone for bacterial expression under T7 promoter. Uses
infusion based cloning method..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7734)
TITLE Cloning Vectors
REFERENCE 2 (bases 1 to 7734)
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"
COMMENT Sequence Label: pET15-MHL
FEATURES Location/Qualifiers
source 1..7734
/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..2153
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2079..2098)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2239..2261
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2255..2446)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3758..4840)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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"
primer_bind 4802..4821
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4841..4918)
/gene="lacI"
/label=lacI promoter
primer_bind 5124..5143
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5227..5246
/label=T7
/note="T7 promoter, forward primer"
promoter 5227..5245
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5246..5270
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
RBS 5285..5307
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5317..5334
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5347..5367
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
protein_bind complement(5395..5428)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter 5443..5888
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS join(5889..5975,5976..7310)
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
primer_bind complement(7461..7479)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 7475..7522
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(join(7697..7702,7703..7719,7720..7725))
/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 tattgaaaaa 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 gcaatggtgc actctcagta caatctgctc tgatgccgca
2101 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
2161 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
2221 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
2281 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
2341 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
2401 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
2461 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
2521 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
2581 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
2641 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
2701 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
2761 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
2821 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
2881 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc
2941 gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt
3001 gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt
3061 tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg
3121 caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca
3181 tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag
3241 gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct
3301 ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat
3361 ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc
3421 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
3481 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
3541 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
3601 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
3661 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
3721 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
3781 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
3841 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
3901 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
3961 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
4021 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
4081 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
4141 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
4201 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
4261 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
4321 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
4381 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
4441 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
4501 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
4561 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
4621 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
4681 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
4741 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
4801 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
4861 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
4921 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
4981 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
5041 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
5101 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
5161 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
5221 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa
5281 taattttgtt taactttaag aaggagatat accatgcatc atcatcatca tcacagcagc
5341 ggcagagaaa acttgtattt ccagggccat atgagttctc ctcctgaaag atccataact
5401 tcgtatagca tacattatac gaagttatgc ggccgcgacg tccacatata cctgccgttc
5461 actattattt agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac
5521 tttatgccca tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt
5581 ttttagttct ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga
5641 ggaaaataga ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat
5701 cgcgcgggtt tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta
5761 tactttggcg tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc
5821 catcttcaaa caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga
5881 catgaacgat gaacatcaaa aagtttgcaa aacaagcaac agtattaacc tttactaccg
5941 cactgctggc aggaggcgca actcaagcgt ttgcgaaaga aacgaaccaa aagccatata
6001 aggaaacata cggcatttcc catattacac gccatgatat gctgcaaatc cctgaacagc
6061 aaaaaaatga aaaatataaa gttcctgagt tcgattcgtc cacaattaaa aatatctctt
6121 ctgcaaaagg cctggacgtt tgggacagct ggccattaca aaacactgac ggcactgtcg
6181 caaactatca cggctaccac atcgtctttg cattagccgg agatcctaaa aatgcggatg
6241 acacatcgat ttacatgttc tatcaaaaag tcggcgaaac ttctattgac agctggaaaa
6301 acgctggccg cgtctttaaa gacagcgaca aattcgatgc aaatgattct atcctaaaag
6361 accaaacaca agaatggtca ggttcagcca catttacatc tgacggaaaa atccgtttat
6421 tctacactga tttctccggt aaacattacg gcaaacaaac actgacaact gcacaagtta
6481 acgtatcagc atcagacagc tctttgaaca tcaacggtgt agaggattat aaatcaatct
6541 ttgacggtga cggaaaaacg tatcaaaatg tacagcagtt catcgatgaa ggcaactaca
6601 gctcaggcga caaccatacg ctgagagatc ctcactacgt agaagataaa ggccacaaat
6661 acttagtatt tgaagcaaac actggaactg aagatggcta ccaaggcgaa gaatctttat
6721 ttaacaaagc atactatggc aaaagcacat cattcttccg tcaagaaagt caaaaacttc
6781 tgcaaagcga taaaaaacgc acggctgagt tagcaaacgg cgctctcggt atgattgagc
6841 taaacgatga ttacacactg aaaaaagtga tgaaaccgct gattgcatct aacacagtaa
6901 cagatgaaat tgaacgcgcg aacgtcttta aaatgaacgg caaatggtac ctgttcactg
6961 actcccgcgg atcaaaaatg acgattgacg gcattacgtc taacgatatt tacatgcttg
7021 gttatgtttc taattcttta actggcccat acaagccgct gaacaaaact ggccttgtgt
7081 taaaaatgga tcttgatcct aacgatgtaa cctttactta ctcacacttc gctgtacctc
7141 aagcgaaagg aaacaatgtc gtgattacaa gctatatgac aaacagagga ttctacgcag
7201 acaaacaatc aacgtttgcg cctagcttcc tgctgaacat caaaggcaag aaaacatctg
7261 ttgtcaaaga cagcatcctt gaacaaggac aattaacagt taacaaataa aaacgcaaaa
7321 gaaaatgccg atatcctatt ggcattgacg tcaggtggca cttttcgagg agatcatgca
7381 catgatgacg aagcttgcgg ccgcactcga ggatccggct gctaacaaag cccgaaagga
7441 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
7501 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg
7561 cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg
7621 atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac
7681 tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg agaa
//
