| 质粒名称: | pOK12 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 49789 |
| 存储实验室: | Joachim Messing |
|
载体骨架基本信息 |
|
| 载体名称: | pUC21 (Addgene plasmid #49787) |
| 载体出品公司: | Messing Lab |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 2135 bp |
| 完整质粒大小: | -- |
| 载体修饰: | lac区和MCS与pUC21相同，KmR基因来自Tn905，ori基因来自pACYC 177。 |
| 载体类型: | 克隆载体 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 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 | lacZ |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13-F20 |
| 3' 测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | pOK12 was a gift from Joachim Messing (Addgene plasmid # 49789 ; http://n2t.net/addgene:49789 ; RRID:Addgene_49789) |
| 参考文献部分: | New pUC-derived cloning vectors with different selectable markers and DNA replication origins. Vieira J, Messing J. Gene. 1991 Apr;100:189-94. 10.1016/0378-1119(91)90365-I PubMed 1905257 |
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 pOK12 2229 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION E. coli cloning vector (KanR, low copy, blue/white selection).
ACCESSION .
VERSION .
KEYWORDS pOK12
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2229)
AUTHORS Vieira J, Messing J
TITLE New pUC-derived cloning vectors with different selectable markers
and DNA replication origins.
JOURNAL Gene. 1991 Apr;100:189-94.
PUBMED 1905257
REFERENCE 2 (bases 1 to 2229)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2229
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(51..595)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(131..150)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(794..813)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 805..1614
/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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(872..891)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
CDS complement(1681..2025)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPLVRGLEIYRCMPWYPGARIRSFCRRVDVIWIRYRRGGRS
RTSGSIPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRP
SQQLRSLNGE"
primer_bind 1806..1828
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1820..1837
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1821..1837
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1844..1863
/label=T7
/note="T7 promoter, forward primer"
promoter 1844..1862
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1877..1893)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1877..1893)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(2021..2037)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2021..2037)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2034..2056)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2045..2061
/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)."
protein_bind 2116..2137
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 gcgtattgga gctttcgcga gggaagtgag agggccgcgg caaagccgtt tttccatagg
61 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
121 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
181 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
241 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
301 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
361 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtttgaa
421 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
481 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
541 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
601 tcatcttatt aaggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
661 catgagatta tcctagagtc gagggggggg ggggaaagcc acgttgtgtc tcaaaatctc
721 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
781 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
841 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
901 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
961 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
1021 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
1081 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
1141 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
1201 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
1261 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
1321 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
1381 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
1441 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
1501 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
1561 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
1621 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
1681 tcattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
1741 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
1801 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
1861 gggcgaattg gggatcgatc cactagttct agagcggccg ccacggcgat atcggatcca
1921 tatgacgtcg acgcgtctgc agaagcttcg aattcgagct cccgggtacc atggcatgca
1981 tcgatagatc tcgaggcctc ggactagtgg cgtaatcatg gtcatagctg tttcctgtgt
2041 gaaattgtta tccgctcaca attccacaca acatacgagc cgcggaagca taaagtgtaa
2101 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
2161 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
2221 aggcggttt
//
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