| 质粒名称: | 2Bc-T MBP_BoMoC(ed)_F753A_6xH |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 185710 |
| 存储实验室: | Kathleen Collins |
|
载体骨架基本信息 |
|
| 载体名称: | pET His6 LIC cloning vector (2Bc-T) |
| 载体出品公司: | Scott Gradia |
| 空载体大小: | 5899 bp |
| 完整质粒大小: | 8416 bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | B. mori truncated R2 RT / BoMoC |
| 物种来源: | Bombyx mori |
| 插入大小: | 2520 bp |
| 基因突变: | Changed F753 to A |
| 启动子: | T7 |
| 标签/融合蛋白: |
MBP (N terminal on insert)
6xHis (C terminal on insert)
|
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | T7 promoter |
| 3’测序引物 | T7 terminator |
|
文献引用方法 |
|
| 材料和方法部分: | 2Bc-T MBP_BoMoC(ed)_F753A_6xH was a gift from Kathleen Collins (Addgene plasmid # 185710 ; http://n2t.net/addgene:185710 ; RRID:Addgene_185710) |
| 参考文献部分: | Separable structural requirements for cDNA synthesis, nontemplated extension, and template jumping by a non-LTR retroelement reverse transcriptase. Pimentel SC, Upton HE, Collins K. J Biol Chem. 2022 Mar;298(3):101624. doi: 10.1016/j.jbc.2022.101624. Epub 2022 Jan 21. 10.1016/j.jbc.2022.101624 PubMed 35065960 |
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2Bc-T MBP_BoMoC(ed)_F753A_6xH载体质粒基本信息
2Bc-T MBP_BoMoC(ed)_F753A_6xH质粒图谱载体图谱和2Bc-T MBP_BoMoC(ed)_F753A_6xH载体序列质粒序列多克隆位点信息
_F753A_6xH质粒图谱.png)
2Bc-T MBP_BoMoC(ed)_F753A_6xH质粒载体简介
2Bc-T MBP_BoMoC(ed)_F753A_6xH质粒序列载体序列
LOCUS 2Bc-T MBP_BoMoC(ed)_F753A_6xH 8416 bp ds-DNA circular SYN 01-AUG-2022
DEFINITION Recombinant B. mori truncated R2 RT expression.
ACCESSION .
VERSION .
KEYWORDS 2Bc-T MBP_BoMoC(ed)_F753A_6xH
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8416)
AUTHORS Pimentel SC, Upton HE, Collins K
TITLE Separable structural requirements for cDNA synthesis, nontemplated
extension, and template jumping by a non-LTR retroelement reverse
transcriptase.
JOURNAL J Biol Chem. 2022 Mar;298(3):101624. doi: 10.1016/j.jbc.2022.101624.
Epub 2022 Jan 21.
PUBMED 35065960
REFERENCE 2 (bases 1 to 8416)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 1, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8416
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 168..187
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 421..438
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 453..592
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(518..537)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 678..700
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(694..885)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 2343..2362
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 2446..2465
/label=T7
/note="T7 promoter, forward primer"
promoter 2446..2464
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 2553..2575
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 6223..6243
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 6250..6267
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6391..6409)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6405..6452
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(6818..6846)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(6873..6891)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6959..7063
/gene="bla"
/label=AmpR promoter
CDS 7064..7924
/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(7282..7301)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8095..267
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
61 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
301 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
361 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
421 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
481 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
541 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
601 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
661 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
721 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
781 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
841 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
901 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
961 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
1021 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
1081 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
1141 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
1201 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
1261 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
1321 caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg
1381 ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg
1441 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
1501 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
1561 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
1621 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg
1681 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
1741 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
1801 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
1861 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
1921 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
1981 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
2041 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
2101 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta
2161 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
2221 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac gcggcctgcc
2281 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
2341 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
2401 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
2461 tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt aattaagcgg
2521 ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa ggagatatag
2581 ttcatgaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
2641 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
2701 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
2761 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
2821 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
2881 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
2941 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
3001 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
3061 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
3121 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
3181 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
3241 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
3301 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
3361 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
3421 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
3481 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc
3541 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
3601 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
3661 gaagccctgg cagccgcgca gactgctgcg gccgcagcta tgggtcgtcg tcgtgcgtgc
3721 cgtgcgatgc gtccgaagac cgcgggtcgt cgtaacgacc tgcacgacga tcgtaccgcg
3781 agcgcgcaca agaccagccg tcagaaacgt cgtgcggagt acgcgcgtgt gcaagaactg
3841 tataagaaat gccgtagccg tgcggcggcg gaagtgatcg atggtgcgtg cggtggcgtt
3901 ggtcacagcc tggaggaaat ggaaacctac tggcgtccga ttctggaacg tgtgagcgac
3961 gcgccgggtc cgaccccgga ggcgctgcac gcgctgggtc gtgcggaatg gcacggtggc
4021 aaccgtgatt atacccagct gtggaagccg atcagcgttg aggaaattaa agcgagccgt
4081 ttcgactggc gtaccagccc gggtccggat ggtatccgta gcggccagtg gcgtgcggtg
4141 ccggttcacc tgaaggcgga aatgttcaac gcgtggatgg cgcgtggcga gatcccggaa
4201 attctgcgtc aatgccgtac cgtgtttgtt ccgaaagttg agcgtccggg tggcccgggt
4261 gaataccgtc cgatcagcat tgcgagcatc ccgctgcgtc acttccacag cattctggcg
4321 cgtcgtctgc tggcgtgctg cccgccggac gcgcgtcagc gtggctttat ctgcgcggat
4381 ggtaccctgg agaacagcgc ggtgctggac gcggttctgg gtgatagccg taagaaactg
4441 cgtgaatgcc acgtggcggt tctggacttc gcgaaggcgt ttgataccgt gagccacgag
4501 gcgctggttg aactgctgcg tctgcgtggc atgccggagc agttctgcgg ttacattgcg
4561 cacctgtatg acaccgcgag caccaccctg gcggtgaaca acgaaatgag cagcccggtg
4621 aaagttggcc gtggtgttcg tcaaggcgac ccgctgagcc cgatcctgtt taacgtggtt
4681 atggatctga ttctggcgag cctgccggag cgtgtgggtt accgtctgga gatggaactg
4741 gttagcgcgc tggcgtatgc ggacgatctg gtgctgctgg cgggcagcaa ggttggtatg
4801 caggaaagca tcagcgcggt ggactgcgtt ggccgtcaaa tgggtctgcg tctgaactgc
4861 cgtaaaagcg cggtgctgag catgatcccg gatggtcacc gtaagaaaca ccactacctg
4921 accgagcgta ccttcaacat tggtggcaag ccgctgcgtc aggtgagctg cgttgaacgt
4981 tggcgttatc tgggcgtgga ctttgaggcg agcggttgcg ttaccctgga acacagcatc
5041 agcagcgcgc tgaacaacat tagccgtgcg ccgctgaaac cgcagcaacg tctggagatc
5101 ctgcgtgcgc acctgattcc gcgtttccag cacggcgccg ttctgggtaa catcagcgac
5161 gatcgtctgc gtatgctgga tgtgcagatt cgtaaggcgg ttggtcaatg gctgcgtctg
5221 ccggcggacg tgccgaaagc gtactatcac gcggcggttc aagatggtgg cctggcgatc
5281 ccgagcgtgc gtgcgaccat cccggacctg attgttcgtc gttttggtgg cctggatagc
5341 agcccgtgga gcgtggcgcg tgcggcggcg aagagcgaca aaattcgtaa gaaactgcgt
5401 tgggcgtgga agcagctgcg tcgtttcagc cgtgtggata gcaccaccca acgtccgagc
5461 gttcgtctgt tttggcgtga gcacctgcac gcgagcgttg acggtcgtga gctgcgtgaa
5521 agcacccgta ccccgaccag caccaaatgg attcgtgaac gttgcgcgca gattaccggt
5581 cgtgatttcg tgcaatttgt tcacacccac atcaacgcgc tgccgagccg tattcgtggc
5641 agccgtggcc gtcgtggtgg cggtgagagc agcctgacct gccgtgcggg ttgcaaagtg
5701 cgtgaaacca ccgcgcacat cctgcagcaa tgccaccgta cccacggcgg tcgtatcctg
5761 cgtcacaaca agattgtgag cttcgttgcg aaggcgatgg aggaaaacaa atggaccgtg
5821 gagctggaac cgcgtctgcg taccagcgtt ggcctgcgta aaccggcgat cattgcgagc
5881 cgtgatggcg tgggtgttat cgtggacgtt caggtggtta gcggtcaacg tagcctggat
5941 gagctgcacc gtgaaaagcg taacaaatac ggcaaccacg gtgagctggt tgagctggtt
6001 gcgggccgtc tgggtctgcc gaaagcggag tgcgtgcgtg cgaccagctg caccattagc
6061 tggcgtggcg tttggagcct gaccagctat aaagagctgc gtagcatcat tggtctgcgt
6121 gaaccgaccc tgcagatcgt gccgattctg gcgctgcgtg gcagccacat gaactggacc
6181 cgttttaacc aaatgaccag cgtgatgggt ggcggtgttg gtgagaacct ctacttccaa
6241 tccggctctc atcaccatca ccatcactaa taaccaactc cataaggatc cgcgatcgcg
6301 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
6361 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
6421 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggacatc
6481 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
6541 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
6601 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
6661 tggacgacat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
6721 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
6781 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
6841 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
6901 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
6961 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
7021 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
7081 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
7141 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
7201 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
7261 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
7321 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
7381 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
7441 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
7501 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
7561 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
7621 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
7681 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
7741 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
7801 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
7861 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
7921 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
7981 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
8041 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
8101 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
8161 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
8221 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
8281 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
8341 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
8401 gcggtcgggc tgaacg
//
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