| 质粒名称: | pET_RTX |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 102787 |
| 存储实验室: | Andrew Ellington |
|
载体骨架基本信息 |
|
| 载体名称: | modified pET21 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 8230 bp |
| 载体修饰: | -- |
| 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: | reverse transcription xenopolymerase,RTX |
| 物种来源: | -- |
| 插入大小: | 2328 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7 promoter + lac operator |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NdeI (销毁后未知) |
| 3' 克隆位点: | BamHI (销毁后未知) |
| 5' 测序引物 | gatctcgatcccgcgaaattaatacg |
| 3' 测序引物 | gctcagcggtggcagc |
|
文献引用方法 |
|
| 材料和方法部分: | pET_RTX was a gift from Andrew Ellington (Addgene plasmid # 102787 ; http://n2t.net/addgene:102787 ; RRID:Addgene_102787) |
| 参考文献部分: | Synthetic evolutionary origin of a proofreading reverse transcriptase. Ellefson JW, Gollihar J, Shroff R, Shivram H, Iyer VR, Ellington AD. Science. 2016 Jun 24;352(6293):1590-3. doi: 10.1126/science.aaf5409. 10.1126/science.aaf5409 PubMed 27339990 |
买家导航
pETRTX载体基本信息
pETRTX载体图谱质粒图谱和多克隆位点信息
pETRTX载体简介
pETRTX载体序列
LOCUS pET_RTX 8230 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses the Reverse Transcriptase gene.
ACCESSION .
VERSION .
KEYWORDS pET_RTX
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8230)
AUTHORS Ellefson JW, Gollihar J, Shroff R, Shivram H, Iyer VR, Ellington AD
TITLE Synthetic evolutionary origin of a proofreading reverse
transcriptase.
JOURNAL Science. 2016 Jun 24;352(6293):1590-3. doi: 10.1126/science.aaf5409.
PUBMED 27339990
REFERENCE 2 (bases 1 to 8230)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8230
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 35..139
/gene="bla"
/label=AmpR promoter
CDS 140..1000
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(358..377)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin complement(1042..1555)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1237..1258)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1449..1468
/label=F1ori-R
/note="F1 origin, reverse primer"
rep_origin 1666..2254
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2155..2174
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2408..2425
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(2447..2466)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2607..2629
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2623..2814)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3371..4453)
/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
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARPRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 4415..4434
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4454..4531)
/gene="lacI"
/label=lacI promoter
terminator 4684..4770
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4867..4953
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5050..5136
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5233..5319
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5416..5502
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 5637..5656
/label=T7
/note="T7 promoter, forward primer"
promoter 5637..5655
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5656..5680
/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 5695..5717
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
RBS 7653..7661
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 8060..8077
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8144..8162)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 8158..8205
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
61 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
301 ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta aagttctgct
361 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
1021 atatatactt tagattgatt taccccggtt gataatcaga aaagccccaa aaacaggaag
1081 attgtataag caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
1141 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
1201 tcaaaagaat agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
1261 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
1321 ctacgtgaac catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
1381 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
1441 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
1501 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag
1561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
1621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
1681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
1741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
1801 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
1861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
1921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
1981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
2041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
2101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
2161 cgcctggtat ctttatagtc ctgttgggtt tcgccacctc tgacttgagc gtcgattttt
2221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
2281 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
2341 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
2401 cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac tctcagtaca atctgctctg
2461 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
2521 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
2581 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
2641 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc
2701 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
2761 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
2821 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
2881 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
2941 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcc
3001 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
3061 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
3121 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
3181 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
3241 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
3301 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
3361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
3421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
3481 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
3541 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
3601 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
3661 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
3721 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
3781 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
3841 gatatttatg ccagccagcc agacgcggac gcgccgagac agaacttaat gggcccgcta
3901 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
3961 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
4021 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
4081 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
4141 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
4201 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
4261 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
4321 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
4381 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
4441 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
4501 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
4561 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
4621 aatggtgcat gccggcatgc cgccctttcg tcttcaagaa ttaattccca attccccagg
4681 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
4741 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
4801 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc
4861 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
4921 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
4981 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattaattc
5041 cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
5101 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
5161 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa
5221 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
5281 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
5341 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
5401 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
5461 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
5521 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
5581 aattggggat cggaattaat tcccggttta aaccggggat ctcgatcccg cgaaattaat
5641 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
5701 taactttaag aaggagatat acatatgatt cttgacacgg attacatcac ggaagacggc
5761 aagccggtta tccgtatttt caagaaagaa aacggcgaat tcaagattga atacgatcgg
5821 acatttgaac cgtacctgta cgctctcctc aaggatgata gcgcaatcga agaagtgaaa
5881 aaaatcaccg cagagcggca tggcacagtg gtaacagtta agcgggtcga gaaagtgcag
5941 aagaagttct taggccggcc agtcgaagta tggaaattat acttcacaca tccacaggac
6001 gttccggcga tcatggataa gattcgggag catccggcgg taatcgatat ctatgaatac
6061 gatattccgt tcgctattcg ctaccttatt gacaaaggtt tagttccaat ggagggtgat
6121 gaggaactta aactgttagc attcgatatc gaaacacttt atcacgaagg tgaagagttt
6181 gccgaaggtc cgattttaat gatctcgtac gccgatgaag aaggcgcacg cgtaattacg
6241 tggaaaaatg tggacctccc gtacgtagac gtagtgagca ctgagcgcga gatgattaaa
6301 cgtttccttc gggtagtaaa agaaaaagac ccagacgtgc tgattacgta taacggcgac
6361 aactttgatt ttgcctatct caagaagcgt tgcgaaaagt taggcattaa tttcgccctg
6421 ggtcgggacg gttcagagcc gaaaattcag cggatgggcg accgctttgc tgtggaggta
6481 aaaggtcgca tccatttcga tttatatccg gttatccggc gcaccatcaa cttgccgact
6541 tacacacttg aagcagttta cgaagcggtg ttcggccaac caaaagaaaa ggtttatgcc
6601 gaggagatta ccaccgcatg ggaaactggc gaaaacttgg agcgggtggc tcggtattcc
6661 atggaagatg ccaaggtgac ctacgaactg ggcaaagagt ttttaccgat ggaagcacaa
6721 ttaagccgcc ttattggtca gtccctctgg gatgtgtcgc gttcttcaac gggcaattta
6781 gtcgaatggt ttcttcttcg gaaagcgtac gagcgtaacg agcttgctcc aaataagcca
6841 gacgaaaaag aattggctcg gcgccatcag tcacatgagg gcggctacat taaggagcca
6901 gaacggggct tgtgggagaa catcgtctac cttgattttc ggtctcttta tccgtctatt
6961 atcatcacac ataacgtctc gccagatacc ctgaaccgtg aaggctgtaa agaatatgat
7021 gtggcaccac aggtcggcca tcgtttttgt aaagacttcc cgggcttcat tccatctctt
7081 ctgggtgatt tgttagaaga gcgtcaaaag atcaagaaac gtatgaaagc gacaattgac
7141 ccaattgaac gcaaattact tgattaccgt cagcgtgcaa tcaagatcct cgcgaactct
7201 ctgtacggtt attacggcta cgcacgcgcc cggtggtatt gcaaagaatg tgcagaatca
7261 gtcattgctt ggggtcggga gtacctgacc atgacgatta aggaaattga ggagaaatac
7321 ggtttcaagg tcatctatag tgacacggat ggtttctttg caacgattcc aggtgcggac
7381 gcagaaactg taaagaaaaa ggcaatggag ttcttgaagt atattaatgc gaagttgcca
7441 ggcgccctgg aattagagta cgaaggtttt tataagcgtg gcctgttcgt gacaaagaag
7501 aaatacgcgg taattgacga ggaaggcaag atcacaactc gtggcttgga aattgttcgt
7561 cgcgattgga gcgagatcgc aaaggagacc caagctcgtg tgttggaggc cctcctgaag
7621 gatggtgacg tcgaaaaagc cgtacgcatc gttaaggagg ttacagagaa gcttagcaag
7681 tatgaggtcc caccagagaa acttgttatt cataaacaaa tcactcgcga ccttaaagac
7741 tataaggcca ctggtccaca cgtcgccgta gcaaagcggc ttgcggctcg gggcgtcaag
7801 attcggccag gcacggttat tagttacatc gtcctcaaag gctcaggccg gattgttgat
7861 cgcgcgattc catttgatga atttgatccg acgaagcata aatatgatgc ggaatattac
7921 attgaaaaac aggttctgcc ggcggtggag cgcatcttac gtgcgttcgg ctatcgcaag
7981 gaggatttgc ggtaccagaa aactcgtcaa gtcggtttga gtgcctggct gaagccgaaa
8041 ggtacctgat aactccagcc atcaccatca ccatcactga taagcggccg cattgatccg
8101 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
8161 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
8221 atatccggat
//
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