| 质粒名称: | pET/ELP-I-CAT |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 71461 |
| 存储实验室: | David Wood |
|
载体骨架基本信息 |
|
| 载体名称: | pET21(+) |
| 载体出品公司: | Novagen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 添加了ELP和intein标签 |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
ELP（（VPGXG）110）（主干上的N终端）
intein（主干上的N终端）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 to sequence ELP |
| 3' 测序引物 | F1ori-R |
|
文献引用方法 |
|
| 材料和方法部分: | pET/ELP-I-CAT was a gift from David Wood (Addgene plasmid # 71461 ; http://n2t.net/addgene:71461 ; RRID:Addgene_71461) |
| 参考文献部分: | Simple bioseparations using self-cleaving elastin-like polypeptide tags. Banki MR, Feng L, Wood DW. Nat Methods. 2005 Sep;2(9):659-61. 10.1038/nmeth787 PubMed 16074986 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pET/ELP-I-CAT 8292 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION ELP-intein purification system.
ACCESSION .
VERSION .
KEYWORDS pET/ELP-I-CAT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8292)
AUTHORS Banki MR, Feng L, Wood DW
TITLE Simple bioseparations using self-cleaving elastin-like polypeptide
tags.
JOURNAL Nat Methods. 2005 Sep;2(9):659-61.
PUBMED 16074986
REFERENCE 2 (bases 1 to 8292)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8292
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(99..118)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 309..330
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 494..598
/gene="bla"
/label=AmpR promoter
CDS 599..1459
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(817..836)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1630..2218
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2119..2138
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2372..2389
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2404..2546
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2472..2491)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2632..2654
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2648..2839)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3648..4730)
/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 4692..4711
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4731..4808)
/gene="lacI"
/label=lacI promoter
primer_bind 5014..5033
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5117..5136
/label=T7
/note="T7 promoter, forward primer"
promoter 5117..5135
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5136..5160
/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 5179..5201
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 6922..6933
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 7450..8109
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(7516..7535)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 8136..8153
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8206..8224)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 8220..8267
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
3301 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
3361 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
3421 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
3481 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
3541 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
3601 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
3721 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
3781 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
3841 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
3901 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
3961 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
4021 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
4081 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
4141 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
4201 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
4261 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
4321 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
4381 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
4441 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
4501 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
4561 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
4621 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
4681 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
4741 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
4801 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
4861 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
4921 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
4981 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
5041 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
5101 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
5161 cctctaggat ccaataattt tgtttaactt taagaaggag atatacatat gggccacggc
5221 gtgggtgttc cgggcgtggg tgttccgggt ggcggtgtgc cgggcgcagg tgttcctggt
5281 gtaggtgtgc cgggtgttgg tgtgccgggt gttggtgtac caggtggcgg tgttccgggt
5341 gcaggcgttc cgggtggcgg tgtgccgggc gtgggtgttc cgggcgtggg tgttccgggt
5401 ggcggtgtgc cgggcgcagg tgttcctggt gtaggtgtgc cgggtgttgg tgtgccgggt
5461 gttggtgtac caggtggcgg tgttccgggt gcaggcgttc cgggtggcgg tgtgccgggc
5521 gtgggtgttc cgggcgtggg tgttccgggt ggcggtgtgc cgggcgcagg tgttcctggt
5581 gtaggtgtgc cgggtgttgg tgtgccgggt gttggtgtac caggtggcgg tgttccgggt
5641 gcaggcgttc cgggtggcgg tgtgccgggc gtgggtgttc cgggcgtggg tgttccgggt
5701 ggcggtgtgc cgggcgcagg tgttcctggt gtaggtgtgc cgggtgttgg tgtgccgggt
5761 gttggtgtac caggtggcgg tgttccgggt gcaggcgttc cgggtggcgg tgtgccgggc
5821 gtgggtgttc cgggcgtggg tgttccgggt ggcggtgtgc cgggcgcagg tgttcctggt
5881 gtaggtgtgc cgggtgttgg tgtgccgggt gttggtgtac caggtggcgg tgttccgggt
5941 gcaggcgttc cgggtggcgg tgtgccgggc gtgggtgttc cgggcgtggg tgttccgggt
6001 ggcggtgtgc cgggcgcagg tgttcctggt gtaggtgtgc cgggtgttgg tgtgccgggt
6061 gttggtgtac caggtggcgg tgttccgggt gcaggcgttc cgggtggcgg tgtgccgggc
6121 gtgggtgttc cgggcgtggg tgttccgggt ggcggtgtgc cgggcgcagg tgttcctggt
6181 gtaggtgtgc cgggtgttgg tgtgccgggt gttggtgtac caggtggcgg tgttccgggt
6241 gcaggcgttc cgggtggcgg tgtgccgggc gtgggtgttc cgggcgtggg tgttccgggt
6301 ggcggtgtgc cgggcgcagg tgttcctggt gtaggtgtgc cgggtgttgg tgtgccgggt
6361 gttggtgtac caggtggcgg tgttccgggt gcaggcgttc cgggtggcgg tgtgccgggc
6421 gtgggtgttc cgggcgtggg tgttccgggt ggcggtgtgc cgggcgcagg tgttcctggt
6481 gtaggtgtgc cgggtgttgg tgtgccgggt gttggtgtac caggtggcgg tgttccgggt
6541 gcaggcgttc cgggtggcgg tgtgccgggc gtgggtgttc cgggcgtggg tgttccgggt
6601 ggcggtgtgc cgggcgcagg tgttcctggt gtaggtgtgc cgggtgttgg tgtgccgggt
6661 gttggtgtac caggtggcgg tgttccgggt gcaggcgttc cgggtggcgg tgtgccgggc
6721 gtgggtgttc cgggcgtggg tgttccgggt ggcggtgtgc cgggcgcagg tgttcctggt
6781 gtaggtgtgc cgggtgttgg tgtgccgggt gttggtgtac caggtggcgg tgttccgggt
6841 gcaggcgttc cgggtggcgg tgtgccgggc gggctggtga gctcgaacaa caacaacaat
6901 aacaataaca acaacctcgg gatcgaggga aggatttcag aattcgccct cgcagagggc
6961 actcggatct tcgatccggt caccggtaca acgcatcgca tcgaggatgt tgtcggtggg
7021 cgcaagccta ttcatgtcgt ggctgctgcc aaggacggaa cgctgcatgc gcggcccgtg
7081 gtgtcctggt tcgaccaggg aacgcgggat gtgatcgggt tgcggatcgc cggtggcgcc
7141 atcctgtggg cgacacccga tcacaaggtg ctgacagagt acggctggcg tgccgccggg
7201 gaactccgca agggagacag ggtggcgcaa ccgcgacgct tcgatggatt cggtgacagt
7261 gcgccgattc cggcgcgcgt gcaggcgctc gcggatgccc tggatgacaa attcctgcac
7321 gacatgctgg cggaagaact ccgctattcc gtgatccgag aagtgctgcc aacgcggcgg
7381 gcacgaacgt tcggcctcga ggtcgaggaa ctgcacaccc tcgtcgccga aggggttgtt
7441 gtacacaaca tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
7501 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
7561 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
7621 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
7681 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
7741 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
7801 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
7861 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
7921 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
7981 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat
8041 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
8101 ggggcgtaat gagcaagctt gcggccgcac tcgagcacca ccaccaccac cactgagatc
8161 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
8221 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
8281 ctatatccgg at
//
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