| 质粒名称: | pET16B.Pfu |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 12509 |
| 存储实验室: | Sung-Hou Kim |
|
载体骨架基本信息 |
|
| 载体名称: | pET16b |
| 载体出品公司: | Novagen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 5711 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Pfu DNA polymerase |
| 物种来源: | -- |
| 插入大小: | 2325 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pET16B.Pfu was a gift from Sung-Hou Kim (Addgene plasmid # 12509 ; http://n2t.net/addgene:12509 ; RRID:Addgene_12509) |
| 参考文献部分: | Purification, crystallization and preliminary X-ray crystallographic analysis of Pyrococcus furiosus DNA polymerase. Goldman S, Kim R, Hung LW, Jancarik J, Kim SH. Acta Crystallogr D Biol Crystallogr. 1998 Sep 1. 54(Pt 5):986-8. 10.1107/S0907444998000353 PubMed 9757114 |
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 pET16B.Pfu 8032 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacterial expression of Pfu DNA polymerase.
ACCESSION .
VERSION .
KEYWORDS pET16B.Pfu
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8032)
AUTHORS Goldman S, Kim R, Hung LW, Jancarik J, Kim SH
TITLE Purification, crystallization and preliminary X-ray crystallographic
analysis of Pyrococcus furiosus DNA polymerase.
JOURNAL Acta Crystallogr D Biol Crystallogr. 1998 Sep 1. 54(Pt 5):986-8.
PUBMED 9757114
REFERENCE 2 (bases 1 to 8032)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8032
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..38
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
terminator 213..260
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 256..274
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(2661..2672)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS complement(2685..2714)
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
RBS 2727..2749
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 2764..2788
/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 complement(2788..2807)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2789..2807)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2891..2910)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 3116..3193
/gene="lacI"
/label=lacI promoter
primer_bind complement(3213..3232)
/label=LacI-R
/note="LacI, reverse primer"
CDS 5587..5778
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(5772..5794)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 5880..6019
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 5935..5954
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(6034..6051)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6205..6793)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6285..6304)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6964..7824)
/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 7587..7606
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7825..7929)
/gene="bla"
/label=AmpR promoter
primer_bind 7997..8015
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
301 cgggctttgt tagcagccgg atcctcgagc taggattttt taatgttaag ccaggaagtt
361 aggccgactt gtcttgtctt ttggtatctg aggtcttcct ttctgtatcc aaatccctcc
421 aatatcctaa gtaccgctgg aagaacctgg ttctcaatgt aatattctgc gtcatacttg
481 tgctttttgg gatcgtattc ctcagctaga attgccctat tgctaattgg accatcgcct
541 ctaagtacta tgtatccaat taccattcct ggctttattt taactccttt agcagctagt
601 ttctttgcag cagctacgtg aggacctatc gccttatact catgtaatgg tcttgttatc
661 tgctcatata ttgcgagctt ctctggtgga atttcataat tggcaagctt ttgtattact
721 tcttttacta ttctcacagc ttcttcaaca tctccgtgtt ttagtattgt ctccaaaact
781 ctagcttgag tttcttttgc aatttcactc caatctctcc taactatctc taaaccacga
841 gtaatgactt ttccttcttc atctattact gcatacctct tcttcgtaac gaagaatccc
901 ctcttataaa acccttcata ttcaagctct agcagtccag ggagctttga atttatgtat
961 tttacaaatt ctagagcctt tatctttatt tcctcacttt ctcctcctgg gatagttgca
1021 tagagaccat cagtgtcaat gtagaggact ttaaatccaa gcttttcttc gagctccttc
1081 catactaact cgatgtactt tcttccccag gcagtaacgc tctcagcaca ctccttacag
1141 taccatcttg cttttgcata gccataatat ccgtagaaag aatttgctaa gagttttatc
1201 gctttttgtc tatagtcaag gagtattttt tctataggat cttgagtttc cttcattttt
1261 gtcttaatct ttcgtctttc ctctaacaaa tgtcccaaga gacttggtat aaaaccaggg
1321 atgtccttgc agaacttgtg gcctacttga ggagcgatat catagttctt gcatccctca
1381 agatttagag tatcgggaga aacattgtgg gtaattataa tcgagggata tagggctcta
1441 aaatctaggt atactatgtt ttcccacaac cccttttctg gctctttaac gaatccacct
1501 gtgtagctct ccctgagcct tctttgatac tcctcttcac ttggcttgtt tggagctact
1561 tcgtttcttt cgtaggcttt cctaagtaag aaccactcta caaggttccc tgtgcttgac
1621 cttgaaacat cccataaagg ttgtccaact aatcttgaaa gctgaatttc cattggaagg
1681 aattctttcc cgagttcata agttgccttt gcatcttcca tcgagtattt ggcaactctc
1741 tcaaggttct ctccactttc ccaggctttt gctatctcgt cggcgtatac cttctccttt
1801 ggctttccaa aaattgcttc atatacagcc tctagtgtgt atgttgggag atttattgtc
1861 cttgttatta catgatacaa gtcgaaatgt attcttccct tgacttctac agccgtcata
1921 tcgcctattc tctgcatctt gggctcgctt ccatctcttc caatggttaa tttaatccca
1981 agtttttctg cccttttcgc taaatgtggg aagtcgaatg agtctccatt ataagtaact
2041 ataatgtcag gatccttctc cctgataatc ctgagaaatc tctttatcat ctctctctcg
2101 cttgatacaa cctcaacgta tggaagatct atgtttttcc aagtaatcac ctttgcttca
2161 ttttcatctg cataactaat cattataatt gggccttttc caaactcttc tccttcgtga
2221 tagagggttt ctatatcgaa ggcaagaatc tttagctctt cttccccctc cattggtatt
2281 aggcctttgt cgatgaggta tctctttgca aatggaatat cgtattcgaa gatgtccaca
2341 actgctggat gttctctaac tttttctcta atagtgggaa catcttgggg atgttccaaa
2401 taaagtttcc acacggtaat aggcttgccg agaaactttt tctcaacctt ctctacatca
2461 acaattctca caatctttcc atgcctttcc cccgttattt tcttaacttc ttcaatcttt
2521 gaatcatccc tgagaagagc gtaaatgtat ggtctaaaag ttctatcatg ctctatctta
2581 aattttccgt tctctttttt gaatagccta ataacaggtt ttccttcttc agttatgtaa
2641 tccacatcta aaatcatatg acgaccttcg atatggccgc tgctgtgatg atgatgatga
2701 tgatgatgat gatggcccat ggtatatctc cttcttaaag ttaaacaaaa ttatttctag
2761 aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt tcgcgggatc
2821 gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
2881 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
2941 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
3001 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
3061 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac
3121 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
3181 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
3241 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
3301 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
3361 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
3421 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
3481 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
3541 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
3601 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
3661 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
3721 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
3781 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
3841 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
3901 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
3961 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
4021 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
4081 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
4141 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
4201 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagt ttcccgactg
4261 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
4321 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
4381 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
4441 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
4501 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
4561 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
4621 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
4681 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
4741 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
4801 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
4861 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
4921 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
4981 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
5041 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
5101 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
5161 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
5221 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
5281 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
5341 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
5401 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
5461 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
5521 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
5581 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
5641 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
5701 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
5761 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
5821 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
5881 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
5941 cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag
6001 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
6061 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
6121 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
6181 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
6241 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
6301 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
6361 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
6421 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
6481 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
6541 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
6601 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
6661 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
6721 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
6781 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
6841 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
6901 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
6961 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
7021 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
7081 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
7141 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
7201 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
7261 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
7321 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
7381 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
7441 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
7501 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
7561 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
7621 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
7681 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
7741 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
7801 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
7861 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
7921 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
7981 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aa
//
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