| 质粒名称: | pETM6 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 49795 |
| 存储实验室: | Mattheos Koffas |
|
载体骨架基本信息 |
|
| 载体名称: | pET-Duet |
| 载体出品公司: | Novagen |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5000 bp |
| 载体修饰: | AvrII was inserted upstream of the T7 promoter, XbaI was inserted between the operator (lacO) and ribosome binding site (RBS), SpeI was between the multiple-cloning site (MCS) and T7 terminator, and NheI was placed downstream of the T7 terminator |
| 载体类型: | 细菌表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 启动子 | T7 |
| 标签/融合蛋白: | S-tag (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 测序引物: |
T7: TAATACGACTCACTATAGGG T7t: GCTAGTTATTGCTCAGCGG |
|
文献引用方法 |
|
| 材料和方法部分: | pETM6 was a gift from Mattheos Koffas (爱迪基因 plasmid # 49795) |
| 参考文献部分: | ePathBrick: a synthetic biology platform for engineering metabolic pathways in E. coli. Xu P, Vansiri A, Bhan N, Koffas MA. ACS Synth Biol. 2012 Jul 20;1(7):256-66. doi: 10.1021/sb300016b. Epub 2012 May 4. 10.1021/sb300016b PubMed 23651248 |
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 pETM6 5203 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION ePathBrick vector for pathway engineering in E. coli..
ACCESSION .
VERSION .
KEYWORDS pETM6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5203)
AUTHORS Xu P, Vansiri A, Bhan N, Koffas MA
TITLE ePathBrick: a synthetic biology platform for engineering metabolic
pathways in E. coli.
JOURNAL ACS Synth Biol. 2012 Jul 20;1(7):256-66. doi: 10.1021/sb300016b.
Epub 2012 May 4.
PUBMED 23651248
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pETM6|
FEATURES Location/Qualifiers
source 1..5203
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3..27
/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 42..64
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 138..182
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(221..239)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 235..282
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 326..781
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(413..432)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 623..644
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(896..1756)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 1519..1538
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1757..1849)
/gene="bla"
/label=AmpR promoter
rep_origin 1930..2518
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2419..2438
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2672..2689
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2704..2846
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2772..2791)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2932..2954
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2948..3139)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3711..4793)
/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 4755..4774
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4794..4871)
/gene="lacI"
/label=lacI promoter
primer_bind 5077..5096
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5187..3
/label=T7
/note="T7 promoter, forward primer"
promoter 5187..2
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
61 gagatataca tatggcagat ctcaattgga tatcggccgg ccacgcgatc gctgacgtcg
121 gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact
181 cgtctactag tcgcagctta attaacctaa actgctgcca ccgctgagca ataactagca
241 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctagcgaa aggaggagtc
301 gactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
721 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
781 tctggcggca cgatggcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
1021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
1081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
1141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
1201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
1261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
1321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
1381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
1441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
1501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
1561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
1621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
1681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
1741 aatgttgaat actcatactc ttcctttttc aatcatgatt gaagcattta tcagggttat
1801 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggtcatgac
1861 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
1921 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
1981 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
2041 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
2101 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
2161 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
2221 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
2281 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
2341 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
2401 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
2461 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
2521 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
2581 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
2641 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
2701 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
2761 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
2821 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
2881 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
2941 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
3001 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
3061 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
3121 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
3181 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
3241 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
3301 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
3361 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
3421 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
3481 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
3541 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgctag
3601 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
3661 gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
3721 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
3781 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
3841 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
3901 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
3961 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
4021 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
4081 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
4141 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
4201 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
4261 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
4321 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
4381 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
4441 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
4501 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
4561 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
4621 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
4681 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
4741 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
4801 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
4861 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
4921 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
4981 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
5041 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
5101 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agcctaggat
5161 cgagatcgat ctcgatcccg cgaaattaat acgactcact ata
//
