| 质粒名称: | pTpal-fdc |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 78288 |
| 存储实验室: | David Nielsen |
|
载体骨架基本信息 |
|
| 载体名称: | pTrc99A |
| 空载体大小: | 4176 bp |
| 完整质粒大小: | 7837 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PAL2 |
| 物种来源: | A. thaliana (mustard weed) |
| 插入大小: | 2154 bp |
| 启动子: | trc |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NcoI(未破坏) |
| 3’克隆位点: | XbaI(未破坏) |
| 5’测序引物 | acacaggaaacagaccatgg |
| 3’测序引物 | TCTAGATTAGCAAATCGGAA |
|
文献引用方法 |
|
| 材料和方法部分: | pTpal-fdc was a gift from David Nielsen (Addgene plasmid # 78288 ; http://n2t.net/addgene:78288 ; RRID:Addgene_78288) |
| 参考文献部分: | Microbial production of the aromatic building-blocks (S)-styrene oxide and (R)-1,2-phenylethanediol from renewable resources. McKenna R, Pugh S, Thompson B, Nielsen DR. Biotechnol J. 2013 Dec;8(12):1465-75. doi: 10.1002/biot.201300035. Epub 2013 Jul 29. 10.1002/biot.201300035 PubMed 23801570 |
买家导航
pTpalfdc载体基本信息
pTpalfdc载体图谱质粒图谱和多克隆位点信息
pTpalfdc载体简介
pTpalfdc载体序列
LOCUS pTpal-fdc 7837 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses A. thaliana PAL2 and S. cerevisiae FDC1 in E. coli.
ACCESSION .
VERSION .
KEYWORDS pTpal-fdc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7837)
AUTHORS McKenna R, Pugh S, Thompson B, Nielsen DR
TITLE Microbial production of the aromatic building-blocks (S)-styrene
oxide and (R)-1,2-phenylethanediol from renewable resources.
JOURNAL Biotechnol J. 2013 Dec;8(12):1465-75. doi: 10.1002/biot.201300035.
Epub 2013 Jul 29.
PUBMED 23801570
REFERENCE 2 (bases 1 to 7837)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7837
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 193..222
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 230..246
/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 235..257
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind complement(4020..4037)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 4190..4276
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4368..4395
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 4415..4506
/gene="bla"
/label=AmpR promoter
CDS 4507..5367
/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(4725..4744)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5538..6126
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6027..6046
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6280..6297
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 6312..6452
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(6378..6397)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6538..6560
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter 6638..6715
/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 6716..7798
/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(6735..6754)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
241 taacaatttc acacaggaaa cagaccatgg gcgggaggta accaatggat caaatcgaag
301 caatgttgtg cggcggagga gagaagacaa aagtggcggt tactacgaag actttggcag
361 atccattgaa ttggggttta gcagcggatc aaatgaaagg aagtcattta gatgaagtga
421 agaagatggt cgaagagtat cgtagaccag tcgtgaatct tggcggagaa acactgacga
481 tcggacaagt tgctgccatc tccaccgtag gaggcagcgt taaggttgag ttagcggaga
541 cttcaagagc cggtgtgaaa gctagcagtg attgggttat ggagagcatg aacaaaggta
601 ctgacagtta cggagtcacc accggctttg gtgctacttc tcaccggaga accaaaaacg
661 gcaccgcatt acaaacagaa ctcattagat ttttgaacgc cggaatattc ggaaacacga
721 aggagacatg tcacacactg ccgcaatccg ccacaagagc cgccatgctc gtcagagtca
781 acactcttct ccaaggatac tccgggatcc gattcgagat cctcgaagcg attacaagtc
841 tcctcaacca caacatctct ccgtcactac ctctccgtgg aaccattacc gcctccggcg
901 atctcgttcc tctctcttac atcgccggac ttctcaccgg ccgtcctaat tccaaagcca
961 ccggtcccga cggtgaatcg ctaaccgcga aagaagcttt tgagaaagcc ggaatcagta
1021 ctggattctt cgatttacaa cctaaggaag gtttagctct cgttaatggc acggcggttg
1081 gatctggaat ggcgtcgatg gttctattcg aagcgaatgt ccaagcggtg ttagcggagg
1141 ttttatcagc gatcttcgcg gaggttatga gcgggaaacc tgagtttacc gatcatctga
1201 ctcatcgttt aaaacatcat cccggacaaa tcgaagcggc ggcgataatg gagcacatac
1261 tcgacggaag ctcatacatg aaattagctc aaaaggttca cgagatggat ccattgcaga
1321 aaccaaaaca agatcgttac gctcttcgta catctcctca atggctaggt cctcaaattg
1381 aagtaatccg tcaagctacg aaatcgatag agcgtgaaat caactccgtt aacgataatc
1441 cgttgatcga tgtttcgagg aacaaggcga ttcacggtgg taacttccaa ggaacaccaa
1501 tcggagtttc tatggataac acgagattgg cgattgctgc gattgggaag ctaatgtttg
1561 ctcaattctc tgagcttgtt aatgatttct acaacaatgg acttccttcg aatctaactg
1621 cttcgagtaa tccaagtttg gattatggat tcaaaggagc agagattgct atggcttctt
1681 attgttctga gcttcaatac ttggctaatc cagtcacaag ccatgttcaa tcagctgagc
1741 aacataatca agatgtgaac tctcttggtt tgatctcgtc tcgtaaaaca tctgaagctg
1801 tggatattct taagctaatg tcaacaacgt tccttgtggg gatatgtcaa gctgttgatt
1861 tgagacattt ggaggagaat ctgagacaaa ctgtgaagaa cacagtttct caagttgcta
1921 agaaagtgtt aaccactgga atcaacggtg agttacatcc gtcaaggttt tgcgagaagg
1981 acttgcttaa ggttgttgat cgtgagcaag tgttcacgta tgtggatgat ccttgtagcg
2041 ctacgtaccc gttgatgcag agactaagac aagttattgt tgatcacgct ttgtccaacg
2101 gtgagactga gaagaatgca gtgacttcga tctttcaaaa gattggagct tttgaagagg
2161 agcttaaggc tgtgcttcca aaggaagttg aagcggctag agcggcttat gggaatggaa
2221 ctgcgccgat tcctaaccgg attaaggaat gtaggtcgta tccgttgtat aggttcgtga
2281 gggaagagct tggaacgaag ttgttgactg gagaaaaggt tgtgtctccg ggagaggagt
2341 ttgataaggt cttcactgct atgtgtgaag gtaaacttat tgatccgttg atggattgtc
2401 tcaaggaatg gaacggagct ccgattccga tttgctaatc tagagtcgac ctgcaggggg
2461 aggaattata tgaggaagct aaatccagct ttagaattta gagactttat ccaggtctta
2521 aaagatgaag atgacttaat cgaaattacc gaagagattg atccaaatct cgaagtaggt
2581 gcaattatga ggaaggccta tgaatcccac ttaccagccc cgttatttaa aaatctcaaa
2641 ggtgcttcga aggatctttt cagcatttta ggttgcccag ccggtttgag aagtaaggag
2701 aaaggagatc atggtagaat tgcccatcat ctggggctcg acccaaaaac aactatcaag
2761 gaaatcatag attatttgct ggagtgtaag gagaaggaac ctctcccccc aatcactgtt
2821 cctgtgtcat ctgcaccttg taaaacacat atactttctg aagaaaaaat acatctacaa
2881 agcctgccaa caccatatct acatgtttca gacggtggca agtacttaca aacgtacgga
2941 atgtggattc ttcaaactcc agataaaaaa tggactaatt ggtcaattgc tagaggtatg
3001 gttgtagatg acaagcatat cactggtctg gtaattaaac cacaacatat tagacaaatt
3061 gctgactctt gggcagcaat tggaaaagca aatgaaattc ctttcgcgtt atgttttggc
3121 gttcccccag cagctatttt agttagttcc atgccaattc ctgaaggtgt ttctgaatcg
3181 gattatgttg gcgcaatctt gggtgagtcg gttccagtag taaaatgtga gaccaacgat
3241 ttaatggttc ctgcaacgag tgagatggta tttgagggta ctttgtcctt aacagataca
3301 catctggaag gcccatttgg tgagatgcat ggatatgttt tcaaaagcca aggtcatcct
3361 tgtccattgt acactgtcaa ggctatgagt tacagagaca atgctattct acctgtttcg
3421 aaccccggtc tttgtacgga tgagacacat accttgattg gttcactagt ggctactgag
3481 gccaaggagc tggctattga atctggcttg ccaattctgg atgcctttat gccttatgag
3541 gctcaggctc tttggcttat cttaaaggtg gatttgaaag ggctgcaagc attgaagaca
3601 acgcctgaag aattttgtaa gaaggtaggt gatatttact ttaggacaaa agttggtttt
3661 atagtccatg aaataatttt ggtggcagat gatatcgaca tatttaactt caaagaagtc
3721 atctgggcct acgttacaag acatacacct gttgcagatc agatggcttt tgatgatgtc
3781 acttcttttc ctttggctcc ctttgtttcg cagtcatcca gaagtaagac tatgaaaggt
3841 ggaaagtgcg ttactaattg catatttaga cagcaatatg agcgcagttt tgactacata
3901 acttgtaatt ttgaaaaggg atatccaaaa ggattagttg acaaagtaaa tgaaaattgg
3961 aaaaggtacg gatataaata aaagcttggc tgttttggcg gatgagagaa gattttcagc
4021 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
4081 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
4141 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
4201 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
4261 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
4321 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
4381 gacggatggc ctttttgcgt ttctacaaac tctttttgtt tatttttcta aatacattca
4441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
4501 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
4561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
4621 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
4681 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
4741 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
4801 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
4861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
4921 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
4981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
5041 acgatgccta cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
5101 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
5161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
5221 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
5281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
5341 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
5401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
5461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
5521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
5581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
5641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
5701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
5761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
5821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
5881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
5941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
6001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
6061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
6121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
6181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
6241 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
6301 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
6361 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
6421 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
6481 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
6541 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca
6601 attcgcgcgc gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac
6661 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
6721 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
6781 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
6841 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
6901 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
6961 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
7021 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
7081 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
7141 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
7201 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
7261 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
7321 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
7381 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
7441 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
7501 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
7561 tatcccgccg tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
7621 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
7681 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
7741 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
7801 gcaacgcaat taatgtgagt tagcgcgaat tgatctg
//
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