| 质粒名称: | p2NIL |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20188 |
| 存储实验室: | Tanya Parish |
|
载体骨架基本信息 |
|
| 载体名称: | p2NIL |
| 载体出品公司: | -- |
| 载体抗性: | -- |
| 空载体大小: | 4753bp |
| 完整质粒大小: | -- |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 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: | p2NIL was a gift from Tanya Parish (Addgene plasmid # 20188 ; http://n2t.net/addgene:20188 ; RRID:Addgene_20188) |
| 参考文献部分: | Use of a flexible cassette method to generate a double unmarked Mycobacterium tuberculosis tlyA plcABC mutant by gene replacement. Parish T, Stoker NG. Microbiology. 2000 Aug . 146 ( Pt 8)():1969-75. 10.1099/00221287-146-8-1969 PubMed 10931901 |
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 p2NIL 4753 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS p2NIL
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4753)
AUTHORS Parish T, Stoker NG
TITLE Use of a flexible cassette method to generate a double unmarked
Mycobacterium tuberculosis tlyA plcABC mutant by gene replacement.
JOURNAL Microbiology. 2000 Aug . 146 ( Pt 8)():1969-75.
PUBMED 10931901
REFERENCE 2 (bases 1 to 4753)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4753
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(31..49)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(32..49)
/label=SP6
/note="SP6 promoter, forward primer"
terminator 273..316
/label=bacterial terminator
/note="putative bacterial transcription terminator"
rep_origin complement(535..963)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(645..666)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 857..876
/label=F1ori-R
/note="F1 origin, reverse primer"
CDS complement(1476..2291)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPWK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKFLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 2199..2218
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2283..2302
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind complement(2609..2631)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 2717..2857
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 2772..2791
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(2872..2889)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3043..3631)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3123..3142)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3802..4590)
/codon_start=1
/gene="bla(M)"
/product="beta-lactamase lacking the signal sequence"
/label=bla(M)
/note="allows cytosolic expression of beta-lactamase"
/translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
FKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATT
LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
primer_bind 4425..4444
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 agcttcacgt ggtcgacgga tcctatgtat tctatagtgt cacctaaatc gtatgtgtat
61 gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt acaagcctaa
121 ttgtgtagca tctggcttac tgaagcagac cctatcatct ctctcgtaaa ctgccgtcag
181 agtcggtttg gttggacgaa ccttctgagt ttctggtaac gccgtcccgc acccggaaat
241 ggtcagcgaa ccaatcagca gggtcatcgc tagaaatcat ccttagcgaa agctaaggat
301 tttttttatc tgaattggta ccgcggccgc ttaattaaga attcccctgt aatccgggca
361 gcgcaacgga acattcatca gtgtaaaaat ggaatcaata aagccctgcg cagcgcgcag
421 ggtcagcctg aatacgcgtt taatgaccag cacagtcgtg atggcaaggt cagaatagcg
481 ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggattttg ttaaaattcg
541 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
601 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
661 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
721 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
781 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
841 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
901 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
961 cgtactatgg ttgctttgac gagcacgtat aacgtgcttt cctcgttaga atcagagcgg
1021 gagctaaaca ggaggccgat taaagggatt ttagacagga acggtacgcc agaatcctga
1081 gaagtgtttt tataatcagt gaggccaccg agcaaaagag tctgtccatc acgcaaatta
1141 accgttgtcg caatacttct ttgattagta ataacatcac ttgcctgagt agaagaactc
1201 aaactatcgg ccttgctggt aatatccaga acaatcctga atcgccccat catccagcca
1261 gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt
1321 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
1381 caactcagca aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg
1441 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
1501 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
1561 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
1621 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
1681 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaaac
1741 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
1801 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
1861 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
1921 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
1981 ttccagggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
2041 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
2101 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
2161 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
2221 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
2281 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
2341 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
2401 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
2461 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
2521 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggctgcc
2581 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
2641 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
2701 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
2761 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
2821 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
2881 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
2941 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
3001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
3061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
3121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
3181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
3241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
3301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
3361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
3421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
3481 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
3541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
3601 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
3661 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
3721 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
3781 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
3841 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
3901 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
3961 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
4021 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
4081 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
4141 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
4201 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
4261 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
4321 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
4381 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
4441 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
4501 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
4561 agcatctttt actttcacca gcgtttctgg gtgacgcaga tcccgcaaga ggcccggcag
4621 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
4681 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
4741 actaccgcat taa
//
