| 质粒名称: | pGDP2 MCR-1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 118404 |
| 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: | MCR-1 |
| 启动子 | pLac |
| 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: | pGDP2 MCR-1 was a gift from Gerard Wright (Addgene plasmid # 118404 ; http://n2t.net/addgene:118404 ; RRID:Addgene_118404) |
| 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 |
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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 pGDP2 MCR-1 4588 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 pLac promoter..
ACCESSION .
VERSION .
KEYWORDS pGDP2 MCR-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4588)
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 4588)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4588
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 367..389
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2028..2045
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 2107..2134
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS complement(2225..3040)
/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 2948..2967
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3032..3051
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 3162..3750
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3651..3670
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3904..3921
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3936..4075
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4001..4020)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4161..4183
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4177..4368)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
61 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
121 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
181 ccagtaaggc aaccccgcca gcctagccgg gagatctgcg gcggcggtgc tcaacggcct
241 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc
301 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggtctaga
361 aataattttg tttaacttta agaaggagat ataccatggt ccagcatact cctgtgtggt
421 accgacgctc ggtcagtccg tttgttcttg tggcgagtgt tgccgttttc ttgaccgcga
481 ccgccaatct tacctttttt gataaaatca gccaaaccta tcccatcgcg gacaatctcg
541 gctttgtgct gacgatcgct gtcgtgctct ttggcgcgat gctactgatc accacgctgt
601 tatcatcgta tcgctatgtg ctaaagcctg tgttgatttt gctattaatc atgggcgcgg
661 tgaccagtta ttttactgac acttatggca cggtctatga tacgaccatg ctccaaaatg
721 ccctacagac cgaccaagcc gagaccaagg atctattaaa cgcagcgttt atcatgcgta
781 tcattggttt gggtgtgcta ccaagtttgc ttgtggcttt tgttaaggtg gattatccga
841 cttggggcaa gggtttgatg cgccgattgg gcttgatcgt ggcaagtctt gcgctgattt
901 tactgcctgt ggtggcgttc agcagtcata tggccagttt ctttcgcgtg cataagccgc
961 tgcgtagcta tgtcaatccg atcatgccaa tctactcggt gggtaagctt gccagtattg
1021 agtataaaaa agccagtgcg ccaaaagata ccatttatca cgccaaagac gcggtacaag
1081 caaccaagcc tgatatgcgt aagccacgcc tagtggtgtt cgtcgtcggt gagacggcac
1141 gcgccgatca tgtcagcttc aatggctatg agcgcgatac tttcccacag cttgccaaga
1201 tcgatggcgt gaccaatttt agcaatgtca catcgtgcgg cacatcgacg gcgtattctg
1261 tgccgtgtat gttcagctat ctgggcgcgg atgagtatga tgtcgatacc gccaaatacc
1321 aagaaaatgt gctggatacg ctggatcgct tgggcgtaag tatcttgtgg cgtgataata
1381 attcggactc aaaaggcgtg atggataagc tgccaaaagc gcaatttgcc gattataaat
1441 ccgcgaccaa caacgccatc tgcaacacca atccttataa cgaatgccgc gatgtcggta
1501 tgctcgttgg cttagatgac tttgtcgctg ccaataacgg caaagatatg ctgatcatgc
1561 tgcaccaaat gggcaatcac gggcctgcgt attttaagcg atatgatgaa aagtttgcca
1621 aattcacgcc agtgtgtgaa ggtaatgagc ttgccaagtg cgaacatcag tccttgatca
1681 atgcttatga caatgccttg cttgccaccg atgatttcat cgctcaaagt atccagtggc
1741 tgcagacgca cagcaatgcc tatgatgtct caatgctgta tgtcagcgat catggcgaaa
1801 gtctgggtga gaacggtgtc tatctacatg gtatgccaaa tgcctttgca ccaaaagaac
1861 agcgcagtgt gcctgcattt ttctggacgg ataagcaaac tggcatcacg ccaatggcaa
1921 ccgataccgt cctgacccat gacgcgatca cgccgacatt attaaagctg tttgatgtca
1981 ccgcggacaa agtcaaagac cgcaccgcat tcatccgctg actcgagcac caccaccacc
2041 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
2101 ctgagcaaaa ggccatccgt caggatggcc ttctggtggc acttttcggg gaaatgtgcg
2161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
2221 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
2281 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
2341 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
2401 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
2461 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
2521 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
2581 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
2641 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
2701 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
2761 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
2821 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
2881 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
2941 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
3001 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
3061 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
3121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
3181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
3241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
3301 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
3361 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
3421 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
3481 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
3541 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
3601 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
3661 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
3721 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
3781 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
3841 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
3901 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
3961 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
4021 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
4081 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
4141 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
4201 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
4261 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
4321 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
4381 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
4441 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
4501 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
4561 taatacagat gtaggtgttc cacagggt
//
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