| 质粒名称: | pB1H2w2-zif268 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 18045 |
| 存储实验室: | Scot Wolfe |
|
载体骨架基本信息 |
|
| 载体名称: | ColE1 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3700 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 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: | zif268 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 267 bp |
| 基因突变: | Zif268 is fused to the omega subunit of RNA polymerase and is to be used as a positive control. |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Kpn1（未销毁） |
| 3' 克隆位点: | Xba1（克隆过程中销毁） |
| 5' 测序引物 | OMG5 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pB1H2w2-zif268 was a gift from Scot Wolfe (Addgene plasmid # 18045 ; http://n2t.net/addgene:18045 ; RRID:Addgene_18045) |
| 参考文献部分: | A systematic characterization of factors that regulate Drosophila segmentation via a bacterial one-hybrid system. Noyes MB, Meng X, Wakabayashi A, Sinha S, Brodsky MH, Wolfe SA. Nucleic Acids Res. 2008 Mar 10. ():. 10.1093/nar/gkn048 PubMed 18332042 |
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pB1H2w2-zif268载体 pB1H2w2zif268质粒基本信息
pB1H2w2-zif268质粒图谱 pB1H2w2zif268载体图谱和pB1H2w2-zif268载体序列 pB1H2w2zif268质粒序列多克隆位点信息
pB1H2w2-zif268质粒 pB1H2w2zif268载体简介
pB1H2w2-zif268质粒序列 pB1H2w2zif268载体序列
LOCUS pB1H2w2-zif268 3966 bp ds-DNA circular SYN 09-JUN-2020
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pB1H2w2-zif268
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3966)
AUTHORS Noyes MB, Meng X, Wakabayashi A, Sinha S, Brodsky MH, Wolfe SA
TITLE A systematic characterization of factors that regulate Drosophila
segmentation via a bacterial one-hybrid system.
JOURNAL Nucleic Acids Res. 2008 Mar 10. ():.
PUBMED 18332042
REFERENCE 2 (bases 1 to 3966)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 9, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3966
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 50..66
/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 55..77
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 306..575
/codon_start=1
/gene="E. coli rpoZ"
/product="DNA-directed RNA polymerase subunit omega"
/label=RNAP omega
/note="promotes RNA polymerase assembly"
/translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGR"
CDS 585..608
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
rep_origin 1027..1482
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1114..1133)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1324..1345
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 1508..1612
/gene="bla"
/label=AmpR promoter
CDS 1613..2473
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(1831..1850)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2644..3232
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3133..3152
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3386..3403
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3418..3558
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3484..3503)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3644..3666
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3660..3851)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 gaattccggg ctttacactt tatgcttccg gctcgcatta tgtgtcgact tgtgagcgga
61 taacaatttc acacaggaaa cagctatgcc tcaacagcag caaatgcaac ctcccaattc
121 aagtgcggac aacaaccctt tgcaacagca atcatcacaa aataccgtac caaacgtcct
181 caaccaaatt aaccaaatct tttctccaga ggagcaacgc agcttattac aagaagccat
241 cgaaacctgc aagaattttg aaaaaacaca attgtaagtc tcggctccaa gaaggagata
301 tacccatggc acgcgtaact gttcaggacg ctgtagagaa aattggtaac cgttttgacc
361 tggtactggt cgccgcgcgt cgcgctcgtc agatgcaggt aggcggaaag gacccgctcg
421 taccggaaga aaacgataaa accactgtaa tcgcgctgcg cgaaatcgaa gaaggtctga
481 tcaacaacca gatcctcgac gttcgcgaac gccaggaaca gcaagagcag gaagccgctg
541 aattacaagc cgttaccgct attgctgaag gtcgtgcggc cgcggactac aaggatgacg
601 acgacaagtt ccggaccggt tccaagacac ccccccatgg tacccgccca tatgcttgcc
661 ctgtcgagtc ctgcgatcgc cgcttttctc gctcggatga gcttacccgc catatccgca
721 tccataccgg tcagaagccc ttccagtgtc gaatctgcat gcgtaacttc agtcgtagtg
781 accaccttac cacccacatc cgcacccaca caggcgagaa gccttttgcc tgtgacattt
841 gtgggaggaa gtttgccagg agtgatgaac gcaagaggca taccaaaatc catttaagac
901 agaaggacgt ctagagacta gaaaaaggcc gacaagtccc gctccgctga agatcctggc
961 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
1021 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
1081 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
1141 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
1201 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
1261 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
1321 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
1381 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
1441 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
1501 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
1561 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
1621 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
1681 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
1741 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
1801 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
1861 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
1921 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
1981 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
2041 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
2101 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
2161 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
2221 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
2281 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
2341 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
2401 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
2461 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
2521 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
2581 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
2641 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
2701 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
2761 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
2821 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
2881 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
2941 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
3001 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
3061 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
3121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
3181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
3241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
3301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
3361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
3421 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
3481 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
3541 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
3601 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
3661 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
3721 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
3781 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
3841 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
3901 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
3961 gaacgg
//
