| 质粒名称: | pFCcGi |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 59324 |
| 存储实验室: | Sophie Helaine, David Holden |
|
载体骨架基本信息 |
|
| 载体名称: | pFPV25.1 |
| 载体出品公司: | Addgene Plasmid #20668 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4764 bp |
| 完整质粒大小: | 7200 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mCherry |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | rpsM |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | mCherry-R |
| 3' 测序引物 | PBAD-R |
|
文献引用方法 |
|
| 材料和方法部分: | pFCcGi was a gift from Sophie Helaine & David Holden (Addgene plasmid # 59324 ; http://n2t.net/addgene:59324 ; RRID:Addgene_59324) |
| 参考文献部分: | Identification of salmonella pathogenicity island-2 type III secretion system effectors involved in intramacrophage replication of S. enterica serovar typhimurium: implications for rational vaccine design. Figueira R, Watson KG, Holden DW, Helaine S. MBio. 2013 Apr 16;4(2):e00065. doi: 10.1128/mBio.00065-13. 10.1128/mBio.00065-13 PubMed 23592259 |
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 pFCcGi 7471 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Inducible expression of GFP and constitutive expression of mCherry.
ACCESSION .
VERSION .
KEYWORDS pFCcGi
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7471)
AUTHORS Figueira R, Watson KG, Holden DW, Helaine S
TITLE Identification of salmonella pathogenicity island-2 type III
secretion system effectors involved in intramacrophage replication
of S. enterica serovar typhimurium: implications for rational
vaccine design.
JOURNAL MBio. 2013 Apr 16;4(2):e00065. doi: 10.1128/mBio.00065-13.
PUBMED 23592259
REFERENCE 2 (bases 1 to 7471)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7471
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 78..788
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(223..241)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 491..510
/label=mCherry-F
/note="mCherry, forward primer"
CDS complement(1050..1928)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 1955..2239
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 2162..2181
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 2317..3033
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(2347..2375)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 2970..2990
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
primer_bind complement(4909..4926)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5080..5668)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5160..5179)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5839..6699)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6462..6481
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6700..6804)
/gene="bla"
/label=AmpR promoter
primer_bind 6872..6890
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 gcaagacccg ttctaaagcc atcctggctg cagcgggtat cgctgaaaat gttaagatcc
61 tgggaggaga tatacatatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
121 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
181 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
241 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
301 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
361 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
421 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
481 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
541 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
601 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
661 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
721 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
781 acaagtagcg gccgcgaaag ctttaatgcg gtagtttatc acagttaaat tgctaacgca
841 gtcaggcacc gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc
901 tggatgctgt aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatcgat
961 gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc
1021 aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact
1081 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
1141 cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
1201 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
1261 acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca
1321 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
1381 caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg
1441 cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc
1501 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
1561 tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag
1621 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
1681 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
1741 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
1801 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
1861 aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct
1921 tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc
1981 agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg
2041 cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa
2101 agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc
2161 tatgccatag catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa
2221 ctctctactg tttctccata cccgtttttt tgggctagca ggaggaattc accatggtac
2281 ccggggatcc tctagattta agaaggagat atacatatga gtaaaggaga agaacttttc
2341 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
2401 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
2461 actactggaa aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtgtt
2521 caatgctttg cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg
2581 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
2641 cgtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
2701 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
2761 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga
2821 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
2881 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
2941 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
3001 attacacatg gcatggatga actatacaaa taaatgtcca gacctgcagg catgcaagct
3061 atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct tttagccgct
3121 ttagcggcct ttccccctac ccgaagggtg ggggcgcgtg tgcagccccg cagggcctgt
3181 ctcggtcgat cattcagccc ggctcatcct tctggcgtgg cggcagaccg aacaaggcgc
3241 ggtcgtggtc gcgttcaagg tacgcatcca ttgccgccat gagccgatcc tccggccact
3301 cgctgctgtt caccttggcc aaaatcatgg cccccaccag caccttgcgc cttgtttcgt
3361 tcttgcgctc ttgctgctgt tcccttgccc gcacccgctg aatttcggca ttgattcgcg
3421 ctcgttgttc ttcgagcttg gccagccgat ccgccgcctt gttgctcccc ttaaccatct
3481 tgacacccca ttgttaatgt gctgtctcgt aggctatcat ggaggcacag cggcggcaat
3541 cccgacccta ctttgtaggg gagggcgcac ttaccggttt ctcttcgaga aactggccta
3601 acggccaccc ttcgggcggt gcgctctccg agggccattg catggagccg aaaagcaaaa
3661 gcaacagcga ggcagcatgg cgatttatca ccttacggcg aaaaccggca gcaggtcggg
3721 cggccaatcg gccagggcca aggccgacta catccagcgc gaaggcaagt atgcccgcga
3781 catggatgaa gtcttgcacg ccgaatccgg gcacatgccg gagttcgtcg agcggcccgc
3841 cgactactgg gatgctgccg acctgtatga acgcgccaat gggcggctgt tcaaggaggt
3901 cgaatttgcc ctgccggtcg agctgaccct cgaccagcag aaggcgctgg cgtccgagtt
3961 cgcccagcac ctgaccggtg ccgagcgcct gccgtatacg ctggccatcc atgccggtgg
4021 cggcgagaac ccgcactgcc acctgatgat ctccgagcgg atcaatgacg gcatcgagcg
4081 gcccgccgct cagtggttca agcggtacaa cggcaagacc ccggagaagg gcggggcaca
4141 gaagaccgaa gcgctcaagc ccaaggcatg gcttgagcag acccgcgagg catgggccga
4201 ccatgccaac cgggcattag agcgggctgg ccacgacgcc cgcattgacc acagaacact
4261 tgaggcgcag ggcatcgagc gcctgcccgg tgttcacctg gggccgaacg tggtggagat
4321 ggaaggccgg ggcatccgca ccgaccgggc agacgtggcc ctgaacatcg acaccgccaa
4381 cgcccagatc atcgacttac aggaataccg ggaggcaata gaccatgaac gcaatcgaca
4441 gagtgaagaa atccagaggc atcaacgagt tagcggagca gatcgaaccg ctggcccaga
4501 gcatggcgac actggccgac gaagcccggc aggtcatgag ccagacccag caggccagcg
4561 aggcgcaggc ggcggagtgg ctgaaagccc agcgccagac aggggcggca tgggtggagc
4621 tggccaaaga gttgcgggag gtagccgccg aggtgagcag cgccgcgcag agcgcccgga
4681 gcgcgtcgcg ggggtggcac tggaagctat ggctaaccgt gatgctggct tccatgatgc
4741 ctacggtggt gctgctgatc gcatcgttgc tcttgctcga cctgacgcca ctgacaaccg
4801 aggacggctc gatctggctg cgcttggtgg cccgatgaag aacgacagga ctttgcaggc
4861 cataggccga cagctcaagg ccatgggctg tgagcgctct tccgcttcct cgctcactga
4921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
5521 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
5641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
5761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
5821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
5881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
5941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
6001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
6061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
6121 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
6181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
6241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
6301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
6361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
6421 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
6481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
6541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
6601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
6661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
6721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
6841 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
6901 tcaagaattc gagctcggta cccggggatc ccggagttca tgcgtgatgc aatgaaagtg
6961 ccgttctact tcggtgggac ctcactgctt atcgttgttg tcgtgattat ggactttatg
7021 gctcaagtgc aaactctgat gatgtccagt cagtatgagt ctgcattgaa gaaggcgaac
7081 ctgaaaggct acggccgtta attggtcgcc tgagaagtta cggagagtaa aaatgaaagt
7141 tcgtgcttcc gtcaagaaat tatgccgtaa ctgcaaaatc gttaagcgtg atggtgtcat
7201 ccgtgtgatt tgcagtgccg agccgaagca taaacagcgc caaggctgat tttttcgcat
7261 atttttcttg caaagttggg ttgagctggc tagattagcc agccaatctt ttgtatgtct
7321 gtacgtttcc atttgagtat cctgaaaacg ggcttttcag catggtacgt acatattaaa
7381 tagtaggagt gcatagtggc ccgtatagca ggcattaaca ttcctgatca gaaacacgcc
7441 gtgatcgcgt taacttcgat ctacggtgtc g
//
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