| 质粒名称: | pXS-sfGFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 117386 |
| 存储实验室: | Karen Guillemin |
|
载体骨架基本信息 |
|
| 载体名称: | pGEN-mcs (ori15A) |
| 完整质粒大小: | 5883bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sfGFP |
| 物种来源: | Synthetic |
| 插入大小: | 717 bp |
| 启动子: | Ptac |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NdeI(未破坏) |
| 3’克隆位点: | NotI(未破坏) |
| 5’测序引物 | ggctggataaagggaactca |
| 3’测序引物 | gcgggctagctcattatttg |
|
文献引用方法 |
|
| 材料和方法部分: |
pXS-sfGFP was a gift from Karen Guillemin (Addgene plasmid # 117386 ; http://n2t.net/addgene:117386 ; RRID:Addgene_117386)
|
| 参考文献部分: | Modernized Tools for Streamlined Genetic Manipulation and Comparative Study of Wild and Diverse Proteobacterial Lineages. Wiles TJ, Wall ES, Schlomann BH, Hay EA, Parthasarathy R, Guillemin K. MBio. 2018 Oct 9;9(5). pii: mBio.01877-18. doi: 10.1128/mBio.01877-18. 10.1128/mBio.01877-18 PubMed 30301859 |
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 pXS-sfGFP 5883 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION pGEN-mcs with a modular sfGFP expression scaffold.
ACCESSION .
VERSION .
KEYWORDS pXS-sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5883)
AUTHORS Wiles TJ, Wall ES, Schlomann BH, Hay EA, Parthasarathy R, Guillemin
K
TITLE Modernized Tools for Streamlined Genetic Manipulation and
Comparative Study of Wild and Diverse Proteobacterial Lineages.
JOURNAL MBio. 2018 Oct 9;9(5). pii: mBio.01877-18. doi:
10.1128/mBio.01877-18.
PUBMED 30301859
REFERENCE 2 (bases 1 to 5883)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5883
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 128..214
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 367..384
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
CDS complement(929..1789)
/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 1552..1571
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1790..1894)
/gene="bla"
/label=AmpR promoter
primer_bind 1962..1980
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4271..4299
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
RBS 4323..4345
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 4353..5069
/codon_start=1
/product="enhanced blue variant of GFP"
/label=EBFP
/note="mammalian codon-optimized"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind 5003..5024
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 5151..5171
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
rep_origin complement(5302..5847)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(5382..5401)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gatctaaaac actaggcccc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
61 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
121 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
181 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
241 tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
301 tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag accgcttctg
361 cgttctgatt taatctgtat caggctgaaa atcttctctc atccgccaaa acagccaagc
421 tggatctggc aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct
481 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat
541 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc
601 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt
661 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt
721 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta
781 cccgtcttac tgtcaaccgg atctaaaaca ctagttaagg gattttggtc atgagattat
841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
1021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
1081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
1141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
1201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
1261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
1321 catggtcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
1381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
1441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
1501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
1561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
1621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
1681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
1741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
1801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
1861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
1921 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
1981 cctttcgtct tctagacaac atcagcaagg agaaaggggc taccggcgaa ccagcagccc
2041 ctttataaag gcgcttcagt agtcagacca gcatcagtcc tgaaaaggcg ggcctgcgcc
2101 cgcctccagg ttgctactta ccggattcgt aagccatgaa agccgccacc tccctgtgtc
2161 cgtctctgta acgaatctcg cacagcgatt ttcgtgtcag ataagtgaat atcaacagtg
2221 tgagacacac gatcaacaca caccagacaa gggaacttcg tggtagtttc atggccttct
2281 tctccttgcg caaagcgcgg taagaggcta tcctgatgtg gactagacat agggatgcct
2341 cgtggtggtt aatgaaaatt aacttactac ggggctatct tctttctgcc acacaacacg
2401 gcaacaaacc accttcacgt catgaggcag aaagcctcaa gcgccgggca catcatagcc
2461 catatacctg cacgctgacc acactcactt tccctgaaaa taatccgctc attcagaccg
2521 ttcacgggaa atccgtgtga ttgttgccgc atcacgctgc ctcccggagt ttgtctcgag
2581 cacttttgtt acccgccaaa caaaacccaa aaacaactca tacccaaccc aataaaacac
2641 caaaacaaga caaataatca ttgattgatg gttgaaatgg ggtaaacttg acaaacaaac
2701 ccacttaaaa cccaaaacat acccaaacac acaccaaaaa aacaccataa ggagttttat
2761 aaatgttggt attcattgat gacggttcaa caaacatcaa actacagtag caggaaagcg
2821 acggaacaat taaacagcac attagcccga acagcttcaa acgcgagtgg gcagtctctt
2881 ttggtgataa aaaggtcttt aactacacac tgaacggcga acagtattca tttgatccaa
2941 tcagcccgga tgctgtagtc acaaccaata tcgcatggca atacagcgac gttaatgtcg
3001 ttgcagtgca tcacgcctta ctgaccagtg gtctgccggt aagcgaagtg gatattgttt
3061 gcacacttcc tctgacagag tattacgaca gaaataacca acccaatacg gaaaatattg
3121 agcgtaagaa agcaaacttc cggaaaaaaa ttacattaaa tggcggggat acattcacaa
3181 taaaagatgt aaaagtcatg cctgaatcta taccggcagg ttatgaagtt ctacaagaac
3241 tggatgagtt agattcttta ttaattatag atctcggggg caccacatta gatatttctc
3301 aggtaatggg gaaattatcg gggatcagta aaatatacgg agactcatct cttggtgtct
3361 ctctggttac atctgcagta aaagatgccc tttctcttgc gagaacaaaa ggaagtagct
3421 atcttgctga cgatataatc attcacagaa aagataataa ctatctgaag caacgaatta
3481 atgatgagaa cgaaatatca atagtcaccg aagcaatgaa tgaagcactt cgtaaacttg
3541 agcaacgtgt attaaatacg ctcaatgaat tttctggtta tactcatgtt atggttatag
3601 gcggtggcgc agaattaata tgcgatgcag taaaaaaaca cacacagatt cgtgatgaac
3661 gttttttcaa aaccaataac tctcaatatg atttagttaa cggtatgtat ctcataggta
3721 attaatgatg gacaagcgca gaaccattgc cttcaaacta aatccagatg taaatcaaac
3781 agataaaatt gtttgtgata cactggacag tatcccgcaa ggggaacgaa gccgccttaa
3841 ccgggccgca ctgacggcag gtctggcctt atacagacaa gatccccgga cccctttcct
3901 tttatgtgag ctgctgacga aagaaaccac attttcagat atcgtgaata tattgagatc
3961 gctatttcca aaagagatgg ccgattttaa ttcttcaata gtcactcaat cctcttcaca
4021 acaagagcaa aaaagtgatg aagagaccaa aaaaaatgcg atgaagctaa taaattaatt
4081 caattattat tgagttccct ttatccacta tcaggctgga taaagggaac tcaatcaagt
4141 tattttctta ccagtcatta cataatcgtt attatgaaat aatcgtttgc actgtctctg
4201 ttattcaggc aatttcaata aaggcacttg ctcacgctct gtcattttct gaaactctca
4261 tgtggaattc ttgacaatta atcatcggct cgtataatgt gtggaggatc cctagaaata
4321 attttgttta actttaagaa ggagatatac atatgagcaa gggcgaggag ctgttcaccg
4381 gggtggtgcc catcctggtc gagctggatg gcgacgtaaa cggccacaag ttcagcgtgc
4441 gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
4501 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
4561 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
4621 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcacctac aagacccgcg
4681 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
4741 tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac agccacaacg
4801 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
4861 acgtcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
4921 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtc ctgagcaaag
4981 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
5041 ctctcggcat ggacgagctg tacaagtgag cggccgcggg ccccatggat cgatagctgg
5101 tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat ggtccttctt
5161 gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa ataatgagct
5221 agcccgccta atgagcgggc ttttttttct cggcctagga gatacttaac agggaagtga
5281 gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat
5341 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
5401 ccctggcggc tcccccgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
5461 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
5521 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
5581 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
5641 ctggtaattg atttagagga gttagtcttg aagccatgcg ccggttaagg ctaaactgaa
5701 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
5761 gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
5821 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc
5881 tag
//
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