| 质粒名称: | p44K |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 45800 |
| 存储实验室: | Luc Ponchon |
|
载体骨架基本信息 |
|
| 载体名称: | p44k |
| 载体出品公司: | -- |
| 完整质粒大小: | 2453 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | -- |
|
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: | -- |
| 克隆方法: | -- |
|
文献引用方法 |
|
| 材料和方法部分: | p44K was a gift from Luc Ponchon (Addgene plasmid # 45800 ; http://n2t.net/addgene:45800 ; RRID:Addgene_45800) |
| 参考文献部分: | Co-expression of RNA-protein complexes in Escherichia coli and applications to RNA biology. Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte S, Tisne C. Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576. Epub 2013 Jun 26. 10.1093/nar/gkt576 PubMed 23804766 |
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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 p44K 2453 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS p44K
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2453)
AUTHORS Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte
S, Tisne C
TITLE Co-expression of RNA-protein complexes in Escherichia coli and
applications to RNA biology.
JOURNAL Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576.
Epub 2013 Jun 26.
PUBMED 23804766
REFERENCE 2 (bases 1 to 2453)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..2453
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 21..125
/gene="bla"
/label=AmpR promoter
CDS 126..986
/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(344..363)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1157..1745
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1646..1665
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1899..1916
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2033..2054
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2069..2099
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2107..2123
/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 2112..2134
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 2188..2205
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 2397..2442
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
ORIGIN
1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagctt tgagaaagcg
1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
2101 gtggaattgt gagcggataa caatttcaca caggaaacac atatgggcca tggcactagt
2161 ggatccctcg aggcggccgc cggtacccat catcaccatc accactaaga attcgggggc
2221 ggaaaggatt cgcggggaca ccacagggag acggccctgc ggtcgacgta gattcggctg
2281 cgacgtccgc aggccgtgtc tctgtggctt cggacgtgtc ctcggacgcg ggttcgattc
2341 ccgccgcctc caccactgca gatccttagc gaaagctaag gatttttttt aagctttagc
2401 ataacccctt ggggcctcta aacgggtctt gaggggtttt tttacgtcag gtg
//
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