| 质粒名称: | pKEN GFP mut2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20409 |
| 存储实验室: | Stanley Falkow, Raphael Valdivia |
|
载体骨架基本信息 |
|
| 载体名称: | pKEN |
| 载体出品公司: | Ezaz-Nikpay et al, 1994 Protein Science |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3531 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | Aequorea victoria |
| 插入大小: | 716 bp |
| 基因突变: | S65A V68L S72A |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XbaI(未破坏) |
| 3' 克隆位点: | PstI (未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | pKEN GFP mut2 was a gift from Stanley Falkow & Raphael Valdivia (Addgene plasmid # 20409 ; http://n2t.net/addgene:20409 ; RRID:Addgene_20409) |
| 参考文献部分: | FACS-optimized mutants of the green fluorescent protein (GFP). Cormack BP, Valdivia RH, Falkow S. Gene. 1996 . 173(1 Spec No):33-8. 10.1016/0378-1119(95)00685-0 PubMed 8707053 |
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 pKEN GFP mut2 6203 bp ds-DNA circular SYN 21-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pKEN GFP mut2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6203)
AUTHORS Cormack BP, Valdivia RH, Falkow S
TITLE FACS-optimized mutants of the green fluorescent protein (GFP).
JOURNAL Gene. 1996 . 173(1 Spec No):33-8.
PUBMED 8707053
REFERENCE 2 (bases 1 to 6203)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 21, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6203
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 355..371
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(383..401)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(408..424)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 565..1020
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(652..671)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 862..883
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1087..1106)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1206..1228
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1266..1284)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1352..1456
/gene="bla"
/label=AmpR promoter
CDS 1457..2317
/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 complement(1675..1694)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2488..3076
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2977..2996
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3230..3247
/label=L4440
/note="L4440 vector, forward primer"
terminator 3405..3432
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 3524..3610
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 3763..3780
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
CDS complement(3841..4557)
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(3884..3904)
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
protein_bind 5960..5976
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5988..6006)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6013..6029)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 gttctttcct gtacataacc ttcgggcatg gcactcttga aaaagtcatg ctgtttcata
61 tgatctgggt atctcgcaaa gcattgaaga ccatacgcga aagtagtgac aagtgttggc
121 catggaacag gtagttttcc agtagtgcaa ataaatttaa gggtaagttt tccgtatgtt
181 gcatcacctt caccctctcc actgacagaa aatttgtgcc cattaacatc accatctaat
241 tcaacaagaa ttgggacaac tccagtgaaa agttcttctc ctttactcat atgtatatct
301 ccttcttaaa tctagaggat ccccgggtac cgagctcgaa ttccacacat tatattgtta
361 tccgctcaca atgtcaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
421 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
481 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
541 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
601 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
661 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
721 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
781 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
841 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
901 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
961 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
1021 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
1081 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
1141 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
1201 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
1261 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
1321 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
1381 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
1441 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
1501 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
1561 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
1621 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
1681 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
1741 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
1801 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
1861 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
1921 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
1981 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
2041 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
2101 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
2161 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
2221 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
2281 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
2341 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
2401 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
2461 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
2521 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
2581 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
2641 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
2701 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
2761 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
2821 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
2881 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
2941 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
3001 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
3061 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
3121 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
3181 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
3241 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
3301 tcattaatgc agttattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
3361 tgaatgtatt tagaaaaata aacaaaagag tttgtagaaa cgcaaaaagg ccatccgtca
3421 ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
3481 ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
3541 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
3601 gttttatttg atgcctggca gttccctact ctcgcatggg gagaccccac actaccatcg
3661 gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg
3721 ccgccaggca aattctgttt tatcagaccg cttctgcgtt ctgatttaat ctgtatcagg
3781 ctgaaaatct tctctcatcc gccaaaacag ccaagcttgc atgcctgcag gtctggacat
3841 ttatttgtat agttcatcca tgccatgtgt aatcccagca gctgttacaa actcaagaag
3901 gaccatgtgg tctctctttt cgttgggatc tttcgaaagg gcagattgtg tggacaggta
3961 atggttgtct ggtaaaagga cagggccatc gccaattgga gtattttgtt gataatggtc
4021 tgctagttga acgcttccat cttcaatgtt gtgtctaatt ttgaagttaa ctttgattcc
4081 attcttttgt ttgtctgcca tgatgtatac attgtgtgag ttatagttgt attccaattt
4141 gtgtccaaga atgtttccat cttctttaaa atcaatacct tttaactcga ttctattaac
4201 aagggtatca ccttcaaact tgacttcagc acgtgtcttg tagttcccgt catctttgaa
4261 aaatatagtt ctttcctgta cataaccttc gggcatggca ctcttgaaaa agtcatgctg
4321 tttcatatga tctgggtatc tcgcaaagca ttgaagacca tacgcgaaag tagtgacaag
4381 tgttggccat ggaacaggta gttttccagt agtgcaaata aatttaaggg taagttttcc
4441 gtatgttgca tcaccttcac cctctccact gacagaaaat ttgtgcccat taacatcacc
4501 atctaattca acaagaattg ggacaactcc agtgaaaagt tcttctcctt tactcatatg
4561 tatatctcct tcttaaatct agaggatccc cgggtaccga gctctgagag atcccctcat
4621 aatttcccca aagcgtaacc atgtgtgaat aaattttgag ctagtagggt tgcagccacg
4681 agtaagtctt cccttgttat tgtgtagcca gaatgccgca aaacttccat gcctaagcga
4741 actgttgaga gtacgtttcg atttctgact gtgttagcct ggaagtgctt gtcccaacct
4801 tgtttctgag catgaacgcc cgcaagccaa catgttagtt gaagcatcag ggcgattagc
4861 agcatgatat caaaacgctc tgagctgctc gttcggctat ggcgtaggcc tagtccgtag
4921 gcaggacttt tcaagtctcg gaaggtttct tcaatctgca ttcgcttcga atagatatta
4981 acaagttgtt tgggtgttcg aatttcaaca ggtaagttag ttgctagaac ccatggctcc
5041 tttgccgacg ctgagtagat tttaggtgac gggtggtgac aatgagtccg tgtcgagcgc
5101 tgattttttc ggcctttaga gcgagattta tacaatagaa tttggcatga gattggattg
5161 cttttagtca gcctcttata gcctaaagtc tttgagtgac tagatgacat atcatgtaag
5221 ttgctgatag gtttccagtt ttccgctcct aggtctgcat attgtacttt tcctcttact
5281 cgacttaacc agtaccaacc cagcttctca acggatttat accatggcac tttaaagcca
5341 gcatcactga caatgagcgg tgtggtgtta ctcggtagaa tgctcgcaag gtcggctaga
5401 aattggtcat gagctttctt tgaacattgc tctgaaagcg ggaacgcttt ctcataaaga
5461 gtaacagaac gaccgtgtag tgcgactgaa gctcgcaata ccataagtcg tttttgctca
5521 cgaatatcag accagtcaac aagtacaatg ggcatcgtat tgcccgaaca gataaagcta
5581 gcatgccaac ggtatacagc gagtcgctct ttgtggaggt gacgattacc taacaatcgg
5641 tcgattcgtt tgatgttatg ttttgttctc gctttggttg gcaggttacg gccaagttcg
5701 gtaagagtga gagttttaca gtcaagtaat gcgtggcaag ccaacgttaa gctgttgagt
5761 cgttttaagt gtaattcggg gcagaattgg taaagagagt cgtgtaaaat atcgagttcg
5821 cacatcttgt tgtctgatta ttgatttttc gcgaaaccat ttgatcatat gacaagatgt
5881 gtatccacct taacttaatg atttttacca aaatcattag gggattcatc agtaccgagc
5941 tcgaattcca cacattatat tgttatccgc tcacaatgtc aattcgccct atagtgagtc
6001 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
6061 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
6121 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt
6181 agcggcgcat taagcgcggc ggg
//
