| 质粒名称: | pCN-mBFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 63847 |
| 存储实验室: | Howard Salis |
|
载体骨架基本信息 |
|
| 载体名称: | pQE-30 |
| 完整质粒大小: | 3432 bp |
| 载体类型: | Bacterial Expression |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | mBFP |
| 物种来源: | Synthetic |
| 插入大小: | 747 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知 (未破坏) |
| 5’测序引物 | pBRforEco |
|
文献引用方法 |
|
| 材料和方法部分: | pCN-mBFP was a gift from Howard Salis (Addgene plasmid # 63847 ; http://n2t.net/addgene:63847 ; RRID:Addgene_63847) |
| 参考文献部分: | Rational design of a synthetic Entner-Doudoroff pathway for improved and controllable NADPH regeneration. Ng CY, Farasat I, Maranas CD, Salis HM. Metab Eng. 2015 May;29:86-96. doi: 10.1016/j.ymben.2015.03.001. Epub 2015 Mar 10. 10.1016/j.ymben.2015.03.001 PubMed 25769287 |
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 pCN-mBFP 3432 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Encodes a NADPH dependent blue fluorescent protein expressed under a
strong promoter and strong RBS without His-tagged. .
ACCESSION .
VERSION .
KEYWORDS pCN-mBFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3432)
AUTHORS Ng CY, Farasat I, Maranas CD, Salis HM
TITLE Rational design of a synthetic Entner-Doudoroff pathway for improved
and controllable NADPH regeneration.
JOURNAL Metab Eng. 2015 May;29:86-96. doi: 10.1016/j.ymben.2015.03.001. Epub
2015 Mar 10.
PUBMED 25769287
REFERENCE 2 (bases 1 to 3432)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3432
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 989..1083
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(1159..1181)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 1267..1407
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 1322..1341
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(1422..1439)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1593..2181)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1673..1692)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2363..3157)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind 3084..3103
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3392..3410
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ttcttgagca cagctaacac cacgtcgtcc ctatctgctg ccctaggtct atgagtggtt
61 gctggataac gacgtctttt ttttgacagc tagctcagtc ctaggtataa tatattcagg
121 gagaccacag gatccacggt ttccctctac aaataatttt gtttaacttt tactagagga
181 attcttaaag acccaggacg atttaaggag gagaaaagac atgcagaatc tgaacggcaa
241 agtggctttc gtgaccggcg gcagccgcgg catcggcgcg gcgatcgtcc gccgcttggc
301 ggcggacggc gccgacatcg cgttcaccta tgtcagcgcc tcgtcgaaaa acgtggccac
361 cgccctggtg caagaactcg aggccaaggg ccgccgcgct cgcgccatcc aggcggactc
421 ggcggatccg gcccaggtgc ggcaggcggt cgagcaggcc atcgtgcaac tggggccggt
481 ggacgtgctg gtgaacaacg ccggcatctt cctggccggc cccttgggcg aggtgacgct
541 ggacgactac gaacgcacga tgaacatcaa tgtgcgcgcg cctttcgtgg ccatccaggc
601 cgcgcaggcc tcgatgccgg acggcggccg catcatcaac atcggcagct gcctggcgga
661 acgcgccggc cgagccgggg taacgctgta tgccgccagc aagtcggcgc tgctgggcat
721 gacgcgcggc ctggcgcgcg acctgggcgc gcgcggcatc accgccaacg tcgtgcaccc
781 gggcccgatc gacaccgaca tgaatcccgc agatggcgaa cgctcgggcg aactggtggc
841 cgtgctgtcc ttgcctcatt acggcgaggt gcgcgacatc gccggcatgg tggctttcct
901 ggccgggccg gatgggcgct acgtgaccgg tgcgagtctg gcggtggacg gcggcttcgc
961 cgcttgaaag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
1021 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
1081 aatccaagct agcttggcga gattttcagg agctaaggtc tagagctgcc tcgcgcgttt
1141 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
1201 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
1261 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
1321 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
1381 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
1441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
1501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
1561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
1621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
1681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
1741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
1801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
1861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
1921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
1981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
2041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
2101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
2161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
2221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
2281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
2341 tggtctgaca gttaccgagc tctcagaaga actcgtcaag aaggcgatag aaggcgatgc
2401 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
2461 caagctcttc agcaatatca cgggtatcca acgctatgtc ctgatagcgg tccgccacac
2521 ccagccggcc acagtcgttg aatccagaaa agcggccatt ttccaccatg atattcggca
2581 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
2641 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
2701 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
2761 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
2821 ctttctcggc aggagcaagg tgggatgaca ggagatcctg ccccggcact tcgcccaata
2881 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
2941 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
3001 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
3061 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
3121 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
3181 cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
3241 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
3301 ttgctgtcca taaaaccgcc cagtctagct atcgggtacc gtgccacctg acgtctaaga
3361 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
3421 tcacctgtcg ac
//
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