| 质粒名称: | pET-19b_MMLV-RT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 153312 |
| 存储实验室: | Drew Endy, Philippa Marrack |
|
载体骨架基本信息 |
|
| 载体名称: | pET-19b |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | MMLV Reverse Transcriptase |
| 物种来源: | MMLV |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | 10x His（主干上的N端子） |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pET-19b_MMLV-RT was a gift from Drew Endy & Philippa Marrack (Addgene plasmid # 153312 ; http://n2t.net/addgene:153312 ; RRID:Addgene_153312) |
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 pET-19b_MMLV-RT 7789 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression of MMLV reverse transcriptase.
ACCESSION .
VERSION .
KEYWORDS pET-19b_MMLV-RT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7789)
TITLE COVID-19 Diagnostic Toolkit Enzymes
REFERENCE 2 (bases 1 to 7789)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7789
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 101..120
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 204..223
/label=T7
/note="T7 promoter, forward primer"
promoter 204..222
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 223..247
/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 262..284
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 297..326
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 342..356
/codon_start=1
/product="enterokinase recognition and cleavage site"
/label=enterokinase site
/translation="DDDDK"
primer_bind complement(2493..2511)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2507..2554
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(2729..2757)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(2784..2802)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2870..2974
/gene="bla"
/label=AmpR promoter
CDS 2975..3835
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3193..3212)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4006..4594
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4495..4514
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4748..4765
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4780..4919
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4845..4864)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5005..5027
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5021..5212)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6524..7606)
/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 7568..7587
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7607..7684)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
61 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
121 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
181 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
241 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggccatc
301 atcatcatca tcatcatcat catcacagca gcggccatat cgacgacgac gacaagcata
361 tgggccagcc gctccaagtg ctcaccctga acatcgaaga cgaacatcgc ttgcacgaaa
421 ccagcaaaga accggatgtt tcactgggtt ccacttggct gtcggacttt ccgcaagcgt
481 gggcagaaac cggcggtatg ggtctggctg tgcgccaagc tccgctgatt atcccactga
541 aggcaactag taccccggta tccattaaac agtacccgat gagccaggag gcccgtctgg
601 gcattaaacc gcatatccag cgtctgctgg accagggtat cttagtcccg tgccagtccc
661 cgtggaacac cccgctgctg ccggttaaaa aaccgggcac caacgactat cgcccggtac
721 aagatttacg tgaagttaac aagcgcgtag aagatatcca tccgaccgtt cccaacccgt
781 acaatctgct ctcaggtccg ccgccttccc accaatggta caccgttctg gacctgaaag
841 atgccttctt ttgcttgcgt ctgcatccga ctagccagcc cctgtttgca ttcgagtggc
901 gtgacccgga aatggggatc agcggccagc tcacctggac ccgtctgccg caaggtttca
961 aaaactcacc taccctgttt aatgaagctc tgcaccgcga tctggcggat ttccgcattc
1021 aacacccgga tctgattctg ctgcagtatg tggacgacct gctgctggcg gctacctccg
1081 aactggattg ccagcagggc acccgtgcct tactgcaaac cctgggtaac ttaggttatc
1141 gtgctagcgc gaaaaaagcc cagatctgtc agaagcaggt taaatacctg ggttacctcc
1201 tgaaagaagg ccagcgctgg ttaaccgaag ctcgcaagga gacggtcatg ggtcagccaa
1261 ccccgaaaac cccgcgccag ctgcgtgagt tcctgggcac cgcgggcttt tgtcgcctgt
1321 ggatcccggg cttcgcggaa atggctgcgc cgctgtaccc gctcaccaaa ccggggaccc
1381 tgttcaactg gggtccggat cagcaaaagg cttaccagga gatcaaacag gcgttgctga
1441 ccgcaccagc tctcggtctg ccggacctga cgaaaccgtt tgaactgttt gtggacgaaa
1501 aacagggtta cgcaaaaggt gttctgaccc agaaactggg tccatggcgt cgcccggtcg
1561 cctacctgag caagaaactg gacccggttg cagctggctg gccgccgtgt ctccgtatgg
1621 tagctgcaat cgctgtgctg accaaagatg cgggaaaact gaccatgggt caaccgttgg
1681 tgatcctggc accgcatgcg gtagaagccc tggtgaaaca gccgccggac cgttggctgt
1741 ccaacgcccg catgacgcac taccaggcgc tgctgctgga taccgatcgt gttcagttcg
1801 gcccggttgt tgcgctgaac ccggcaactc tgcttccgtt accggaagag ggtctgcagc
1861 acaactgcct tgatatcctc gcagaagcgc acggcacccg tcctgacctg acggaccagc
1921 cactgccgga cgcagatcat acctggtaca ccgacggcag cagcctgctg caagagggcc
1981 aacgcaaagc gggggcagcc gtaaccactg agaccgaggt tatttgggct aaagctctgc
2041 cagccggcac ttctgcacag cgtgcggaac tgatcgctct gacccaagcc ctgaaaatgg
2101 ctgaaggcaa gaaattgaac gtttacaccg acagccgcta tgcgttcgcg actgcccaca
2161 tccacggtga aatctaccgc cgccgcggtt ggctgacgtc gaagggcaaa gaaatcaaaa
2221 acaaagatga aatcttagct ctgctgaaag ctctgtttct gccgaaacgt ctgtccatta
2281 ttcactgccc gggtcaccag aaagggcact ctgcagaagc tcgtggaaac cgtatggcag
2341 accaggctgc gcgtaaagca gccatcactg aaaccccgga tacgtcaact ctgctgattg
2401 aaaacagcag cccgaacagc cgcttgatta actgactcga ggtgatccgg ctgctaacaa
2461 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
2521 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
2581 tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg
2641 acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt
2701 tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca
2761 tgagaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
2821 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
2881 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
2941 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
3001 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
3061 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
3121 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
3181 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
3241 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
3301 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
3361 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
3421 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
3481 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
3541 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
3601 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
3661 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
3721 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
3781 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
3841 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
3901 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
3961 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
4021 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
4081 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
4141 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
4201 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
4261 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
4321 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
4381 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
4441 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
4501 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
4561 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
4621 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
4681 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
4741 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
4801 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
4861 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
4921 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
4981 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
5041 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
5101 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
5161 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
5221 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
5281 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
5341 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
5401 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
5461 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
5521 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
5581 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
5641 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg
5701 agatgcgccg cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt
5761 tggtttgcgc attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga
5821 atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc
5881 gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc
5941 gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga
6001 agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac
6061 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
6121 cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc
6181 gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
6241 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
6301 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
6361 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
6421 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
6481 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
6541 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
6601 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
6661 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
6721 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
6781 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
6841 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
6901 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
6961 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
7021 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
7081 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
7141 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
7201 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
7261 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
7321 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
7381 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
7441 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
7501 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
7561 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
7621 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
7681 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
7741 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagg
//
