| 质粒名称: | pGDP1 NDM-1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 112883 |
| 存储实验室: | Gerard Wright |
|
载体骨架基本信息 |
|
| 载体名称: | pBR322 |
| 载体抗性: | 卡那霉素 |
| 载体类型: | 细菌表达 |
| 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: | NDM-1 |
| 物种来源: | Klebsiella pneumoniae |
| 启动子 | Pbla |
| 标签/融合蛋白: | 6xHis（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 测序引物 | CATGAGACAATAACCCTG |
| 3' 测序引物 | CCAACTCAGCTTCCTTTC |
|
文献引用方法 |
|
| 材料和方法部分: | pGDP1 NDM-1 was a gift from Gerard Wright (Addgene plasmid # 112883 ; http://n2t.net/addgene:112883 ; RRID:Addgene_112883) |
| 参考文献部分: | A Common Platform for Antibiotic Dereplication and Adjuvant Discovery. Cox G, Sieron A, King AM, De Pascale G, Pawlowski AC, Koteva K, Wright GD. Cell Chem Biol. 2017 Jan 19;24(1):98-109. doi: 10.1016/j.chembiol.2016.11.011. Epub 2016 Dec 22. 10.1016/j.chembiol.2016.11.011 PubMed 28017602 |
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 pGDP1 NDM-1 3922 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION This plasmid is part of the Minimal Antibiotic Resistance Platform
(ARP) Kit (Addgene #1000000143). Low copy-number plasmid that
constitutively expresses genes using the Pbla promoter..
ACCESSION .
VERSION .
KEYWORDS pGDP1 NDM-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3922)
AUTHORS Cox G, Sieron A, King AM, De Pascale G, Pawlowski AC, Koteva K,
Wright GD
TITLE A Common Platform for Antibiotic Dereplication and Adjuvant
Discovery.
JOURNAL Cell Chem Biol. 2017 Jan 19;24(1):98-109. doi:
10.1016/j.chembiol.2016.11.011. Epub 2016 Dec 22.
PUBMED 28017602
REFERENCE 2 (bases 1 to 3922)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3922
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 232..254
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 273..290
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 300..317
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 1180..1197
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 1259..1286
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS complement(1377..2192)
/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
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 2100..2119
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2184..2203
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 2314..2902
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2803..2822
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3056..3073
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3088..3227
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3153..3172)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3313..3335
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3329..3520)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 agtaaggcaa ccccgccagc ctagccggga gatctggcct attggttaaa aaatgagctg
61 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
181 tgtatccgct catgagacaa taaccctgat aaatgcttct ctagaaataa ttttgtttaa
241 ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc
301 tggtgccgcg cggcagccat atggaactgc cgaacattat gcacccggtt gcgaaactga
361 gcacggccct ggcagcagcc ctgatgctga gcggttgtat gccgggtgaa attcgtccga
421 ccatcggcca gcaaatggaa acgggcgatc agcgttttgg tgacctggtg ttccgtcagc
481 tggcaccgaa cgtttggcaa catacctcat atctggatat gccgggcttt ggtgcagttg
541 cttcgaatgg tctgatcgtt cgtgatggtg gtcgtgtcct ggtggttgac accgcatgga
601 cggatgacca gaccgcccaa attctgaact ggatcaaaca ggaaattaat ctgccggtcg
661 cactggcagt cgtgacccat gcacaccaag ataaaatggg tggtatggac gcactgcatg
721 cagcaggcat tgctacgtat gcaaacgctc tgtccaatca gctggcgccg caagaaggta
781 tggtggcagc tcagcacagc ctgacctttg cagcaaacgg ttgggtcgaa ccggcaacgg
841 ccccgaattt tggtccgctg aaagtgttct acccgggccc gggtcacacc agtgataaca
901 tcacggttgg cattgatggt accgacatcg ccttcggcgg ttgcctgatt aaagatagta
961 aagcgaaatc cctgggcaat ctgggtgatg ccgacacgga acattacgca gctagcgcac
1021 gtgcttttgg tgcagcattc ccgaaagcat ctatgatcgt gatgagccac tctgcaccgg
1081 attcacgcgc tgcgattacc cacaccgccc gtatggctga caaactgcgt tgaggatccg
1141 aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccactga
1201 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
1261 aaggccatcc gtcaggatgg ccttctggtg gcacttttcg gggaaatgtg cgcggaaccc
1321 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
1381 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
1441 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
1501 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
1561 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
1621 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
1681 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
1741 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
1801 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
1861 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
1921 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
1981 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
2041 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
2101 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
2161 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
2221 gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
2281 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
2341 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
2401 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
2461 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
2521 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
2581 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
2641 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
2701 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
2761 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
2821 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
2881 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
2941 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
3001 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
3061 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
3121 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
3181 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
3241 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
3301 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
3361 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
3421 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
3481 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
3541 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
3601 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
3661 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
3721 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
3781 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
3841 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
3901 tcggtgattc attctgctaa cc
//
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