| 质粒名称: | pRSM3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 49799 |
| 存储实验室: | Mattheos Koffas |
|
载体骨架基本信息 |
|
| 载体名称: | pRSF-Duet |
| 载体出品公司: | Novagen |
| 载体抗性: | 卡那霉素 |
| 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 |
|
文献引用方法 |
|
| 材料和方法部分: | pRSM3 was a gift from Mattheos Koffas (爱迪基因 plasmid # 49799) |
| 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 pRSM3 3777 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION ePathBrick vector for pathway engineering in E. coli..
ACCESSION .
VERSION .
KEYWORDS pRSM3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3777)
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|pRSM3|
FEATURES Location/Qualifiers
source 1..3777
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 520..539
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(559..636)
/gene="lacI"
/label=lacI promoter
primer_bind 842..861
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 952..971
/label=T7
/note="T7 promoter, forward primer"
promoter 952..970
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 971..995
/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 1010..1032
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1106..1150
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(1189..1207)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1203..1250
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1385..2200)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 2108..2127
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(2201..2292)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2308..3057)
/direction=LEFT
/label=RSF ori
/note="Plasmids containing the RSF 1030 origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
primer_bind complement(2318..2335)
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(2558..2577)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3253..558)
/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"
ORIGIN
1 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
61 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
121 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
181 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
241 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
301 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
361 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
421 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
481 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
541 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
601 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
661 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
721 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
781 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
841 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
901 cgatgcgtcc ggcgtagcct aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac
961 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
1021 ttaagaagga gatatacata tggcagatct caattggata tcggccggcc acgcgatcgc
1081 tgacgtcggt accctcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca
1141 catggactcg tctactagtc gcagcttaat taacctaaac tgctgccacc gctgagcaat
1201 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctagcgaaag
1261 gaggagtcga caagctgacg accgggtctc cgcaagtggc acttttcggg gaaatgtgcg
1321 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
1381 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
1441 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
1501 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
1561 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
1621 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
1681 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
1741 attgcgcctg agcgagacga aatacgcggt cgctgttaaa aggacaatta caaacaggaa
1801 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
1861 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
1921 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
1981 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
2041 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
2101 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
2161 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat actcttcctt tttcaatatt
2221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
2281 aaaataaaca aataggcatg cagcgctctt ccgcttcctc gctcactgac tcgctacgct
2341 cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa agcggtaata cggttatcca
2401 cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa aagcaaagca ccggaagaag
2461 ccaacgccgc aggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
2521 cgctcaagcc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
2581 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
2641 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgttggta tctcagttcg
2701 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
2761 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
2821 ctggcagcag ccattggtaa ctgatttaga ggactttgtc ttgaagttat gcacctgtta
2881 aggctaaact gaaagaacag attttggtga gtgcggtcct ccaacccact taccttggtt
2941 caaagagttg gtagctcagc gaaccttgag aaaaccaccg ttggtagcgg tggtttttct
3001 ttatttatga gatgatgaat caatcggtct atcaagtcaa cgaacagcta ttccgttact
3061 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
3121 taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg
3181 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
3241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
3301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
3361 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
3421 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
3481 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
3541 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
3601 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
3661 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
3721 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcc
//
