| 质粒名称: | pTrc-trGPPS(CO)-LS |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 50603 |
| 存储实验室: | Jay Keasling |
|
载体骨架基本信息 |
|
| 载体名称: | E. coli DH10B |
| 载体出品公司: | -- |
| 完整质粒大小: | 6704 bp |
| 载体类型: | 细菌表达，合成生物学 |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称1: | Limonene Synthase |
| 插入大小1: | 1634 bp |
| 基因突变1: | 截短密码子优化的香叶基焦磷酸合成酶 |
| 启动子1: | Trc |
| 5’测序引物1 | GTCAGAATTAACCCGGGGA |
| 3’测序引物1 | CCTGCAGGTCGACTCACTC |
| 插入基因名称2: | ApR |
| 插入大小2: | 860 bp |
| 启动子2: | Trc |
| 5’测序引物2 | ATTTGAACGTTGCGAAGCA |
| 3’测序引物2 | TTTGCAAGCAGCAGATTACG |
| 插入基因名称3: | LacIq |
| 插入大小3: | 1186 bp |
| 启动子3: | Trc |
| 5’测序引物3 | GAGTCAGTGAGCGAGGAAGC |
| 3’测序引物3 | NA |
|
文献引用方法 |
|
| 材料和方法部分: | pTrc-trGPPS(CO)-LS was a gift from Jay Keasling (Addgene plasmid # 50603 ; http://n2t.net/addgene:50603 ; RRID:Addgene_50603) |
| 参考文献部分: | Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production. Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold CJ, Lee TS. Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004. 10.1016/j.ymben.2013.05.004 PubMed 23727191 |
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
-LS质粒图谱.png)
载体简介
载体序列
LOCUS pTrc-trGPPS(CO)-LS 6704 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION pTrc-trGPPS(CO)-LS: A plasmid that has truncated codon optimized
geranylpyrophosphate synthase and truncated limonene synthase.
ACCESSION .
VERSION .
KEYWORDS pTrc-trGPPS(CO)-LS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6704)
AUTHORS Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold
CJ, Lee TS
TITLE Metabolic engineering of Escherichia coli for limonene and perillyl
alcohol production.
JOURNAL Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004.
PUBMED 23727191
REFERENCE 2 (bases 1 to 6704)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6704
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 196..225
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 233..249
/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)."
primer_bind 238..260
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind complement(2890..2907)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 3060..3146
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3238..3265
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3285..3376
/gene="bla"
/label=AmpR promoter
CDS 3377..4237
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3595..3614)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4408..4996
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4897..4916
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5150..5167
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 5182..5322
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(5248..5267)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5408..5430
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter 5508..5585
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 5586..6668
/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
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(5605..5624)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 ctggtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
61 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
121 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
181 ttctgaaatg agctgttgac aattaatcat ccggctcgta taatgtgtgg aattgtgagc
241 ggataacaat ttcacacagg aaacagacca tggaatttga cttcaacaaa tacatggact
301 ccaaagcgat gacggtaaat gaagcactga acaaagcgat ccctctgcgt tatccgcaga
361 aaatctacga aagcatgcgt tacagcctgc tggcaggcgg caagcgtgtt cgtccggttc
421 tgtgtattgc cgcatgtgaa ctggtaggtg gtaccgaaga actggcgatc ccgaccgcgt
481 gcgcaattga aatgatccac acgatgtccc tgatgcacga tgatctgccg tgtatcgaca
541 acgacgatct gcgtcgcggt aaaccgacta accacaaaat tttcggtgag gataccgcag
601 tgactgctgg taacgcactg cactcttacg ccttcgagca tatcgcggtt tctacttcta
661 aaaccgttgg tgctgaccgc atcctgcgta tggtgtccga gctgggtcgt gctactggct
721 ctgaaggtgt tatgggtggt cagatggtag acatcgcatc cgaaggcgat ccgtctatcg
781 acctgcagac cctggaatgg attcacatcc acaaaaccgc aatgctgctg gaatgctccg
841 ttgtttgcgg tgcaatcatt ggcggtgcca gcgaaatcgt aatcgaacgt gcccgtcgct
901 acgcccgctg tgttggtctg ctgttccagg tagttgatga cattctggac gtaactaaaa
961 gcagcgacga actgggtaag actgcgggca aggacctgat ctctgataaa gccacctacc
1021 caaagctgat gggtctggaa aaggccaagg agttctccga tgaactgctg aaccgtgcga
1081 agggtgaact gtcctgcttc gacccagtta aagccgctcc gctgctgggc ctggcagact
1141 acgtggcatt tcgtcagaat taacccgggg atcccgtttt aagaaggaga tatacatgcg
1201 tcgcagtggt aattacaacc cgtcacgttg ggatgtgaat tttatccaat ccctgctgag
1261 cgattacaaa gaggacaaac acgtcattcg ggcaagcgaa ctggttacac tggttaaaat
1321 ggaactggaa aaagaaacag accaaattcg ccaattagag ctgattgacg acttacagcg
1381 catggggctg agcgatcact ttcagaatga gttcaaagag attctgtcat ctatctatct
1441 ggaccaccat tactataaaa acccttttcc gaaagaggaa cgtgatctgt atagcacaag
1501 cctggccttt cggttactgc gtgagcatgg cttccaagta gcacaagagg tattcgatag
1561 cttcaagaat gaagagggcg agttcaaaga atcactgtcc gatgatactc gtggcttatt
1621 acaactgtat gaagcgagct ttctgctgac cgagggggag acgactctgg aaagcgcacg
1681 ggaatttgcg accaaattct tagaagagaa agtaaatgaa ggcggcgtag atggcgatct
1741 gttaacccgc atcgcgtatt ctctggacat tccgttacac tggcgtatta aacggccaaa
1801 cgcgccggtg tggattgagt ggtatcgcaa acgtccagac atgaacccgg tggtgctgga
1861 attagcaatt ttagacttaa acatcgtgca ggctcaattt caggaagaac tgaaagaatc
1921 ttttcgctgg tggcgcaaca ccggcttcgt tgaaaaatta ccgtttgcgc gcgatcgtct
1981 ggttgaatgt tatttctgga ataccggcat tatcgaaccg cgccagcacg cgagcgcccg
2041 tattatgatg ggtaaggtca atgcgctgat tactgtcatt gacgatattt acgatgtata
2101 cggcactctg gaagaactgg agcagtttac agatttaatc cgccggtggg acatcaatag
2161 catcgaccag ctgccagatt acatgcagct gtgttttctg gcgctgaata actttgtgga
2221 cgacacctct tatgacgtca tgaaagaaaa aggcgtgaat gttatccctt atctgcgtca
2281 gagttgggtg gatctggccg ataaatatat ggttgaggca cgctggttct acggtgggca
2341 caaaccgagt ttagaagaat atctggagaa ctcatggcag agtatcagtg ggccatgcat
2401 gctgacgcat atcttctttc gcgtcacgga ttcatttacc aaggagacgg tcgattcctt
2461 atataaatac catgacctgg ttcgttggtc atctttcgtc ctgcggctgg cggacgatct
2521 gggtacgtct gtggaagagg tgtctcgtgg ggatgttccg aaatccttac agtgctacat
2581 gtccgattat aacgcaagtg aagctgaagc tcgcaaacat gtgaagtggc tgatcgccga
2641 agtgtggaag aagatgaacg ctgaacgtgt ttccaaggat agtccgtttg gtaaggattt
2701 tattggttgc gccgttgatc tgggtcgtat ggcccagctg atgtatcata acggtgatgg
2761 tcatgggacc cagcatccta ttattcatca gcagatgacg cgtaccctgt ttgaaccttt
2821 cgccctcgag tgagtcgacc tgcaggcatg caagcttggc tgttttggcg gatgagagaa
2881 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
2941 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
3001 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
3061 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
3121 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
3181 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
3241 aggccatcct gacggatggc ctttttgcgt ttctacaaac tctttttgtt tatttttcta
3301 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
3361 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
3421 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
3481 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
3541 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
3601 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
3661 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
3721 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
3781 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
3841 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
3901 gcgtgacacc acgatgccta cagcaatggc aacaacgttg cgcaaactat taactggcga
3961 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
4021 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
4081 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
4141 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
4201 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
4261 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
4321 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
4381 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
4441 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
4501 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
4561 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
4621 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
4681 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
4741 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
4801 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
4861 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
4921 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
4981 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
5041 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
5101 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
5161 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
5221 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
5281 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
5341 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
5401 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
5461 cagcagatca attcgcgcgc gaaggcgaag cggcatgcat ttacgttgac accatcgaat
5521 ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
5581 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
5641 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
5701 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
5761 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
5821 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
5881 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
5941 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
6001 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
6061 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
6121 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
6181 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
6241 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
6301 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
6361 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
6421 gctcatgtta tatcccgccg tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa
6481 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
6541 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
6601 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
6661 ggcagtgagc gcaacgcaat taatgtgagt tagcgcgaat tgat
//
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