| 质粒名称: | pLXIN2-EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 99204 |
| 存储实验室: | Saroj Mathupala |
|
载体骨架基本信息 |
|
| 载体名称: |
pLXIN2
|
| 载体出品公司: | Mathupala, S.P. |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 6059 bp |
| 完整质粒大小: | 6780 bp |
| 载体修饰: | 将增强型绿色荧光蛋白（egfp）cDNA插入载体中MCS的Ecori和Xhoi位点之间。 |
| 载体类型: | 哺乳动物表达，逆转录病毒；双顺反转录病毒表达载体 |
| 筛选标记: | 新霉素（用G418选择） |
| 高拷贝/低拷贝: | 高拷贝 |
| 宿生菌: | NEB Stable |
| 生长说明： | 必要时可以在大肠杆菌dh5alpha中繁殖。然而，对于维持来说，最好是在重组缺陷菌株中繁殖，如Sure或Stbl。 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EGFP |
| 备用名称： | 增强型绿色荧光蛋白 |
| 物种来源: | 合成纤维；维多利亚绿松花 |
| 插入大小: | 721bp |
| 基因突变: | 为了提高翻译效率，将Kozak序列插入ATG中5'。从egfp-n1载体（genbank u55762）中扩增egfp。 |
| GenBank编号 | U55762 U55762 |
| 克隆方法: | 限制性酶 |
| 5' 克隆位点: | Ecori（未销毁） |
| 3' 克隆位点: | XHOI（未销毁） |
| 5' 测序引物 | CCCTTGAACCTCCTCGTTCGACC |
| 3' 测序引物 | GTGCAATCCATCTTGTTCAATGGC |
|
文献引用方法 |
|
| 材料和方法部分: | pLXIN2-EGFP was a gift from Saroj Mathupala (Addgene plasmid # 99204 ; http://n2t.net/addgene:99204 ; RRID:Addgene_99204) |
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 pLXIN2-EGFP 6777 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION EGFP expressing bicistronic retroviral vector.
ACCESSION .
VERSION .
KEYWORDS pLXIN2-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6777)
TITLE pLXIN2
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6777)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6777
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 144..165
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 223..794
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 244..796
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(388..405)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 615..634
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 796..1599
/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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(859..878)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1469..1488
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
LTR 1659..2252
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(2355..2377)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 2463..2603
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 2518..2537
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(2618..2635)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2789..3377)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2869..2888)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3548..4408)
/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 4171..4190
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4409..4513)
/gene="bla"
/label=AmpR promoter
primer_bind 4581..4599
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
LTR 5201..5376
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 5439..5638
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 5839..6255
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 6185..6207
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 6223..6239
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
CDS 6268..210
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6313..6334)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(6574..6593)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
ORIGIN
1 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
61 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
121 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
181 actctcggca tggacgagct gtacaagtaa ctcgaggatc cgcccctctc cctccccccc
241 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
301 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
361 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
421 tcgtgaagga agcagttcct ctggaagctt cttgaagaca agcaacgtct gtagcgaccc
481 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
541 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt aggatagttg
601 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
661 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
721 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
781 aaacacgatg ataatatggg atcggccatt gaacaagatg gattgcacgc aggttctccg
841 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
901 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
961 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
1021 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
1081 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
1141 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
1201 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
1261 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
1321 aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc
1381 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
1441 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
1501 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
1561 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgata
1621 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
1681 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga
1741 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg
1801 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
1861 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
1921 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
1981 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
2041 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
2101 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
2161 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
2221 gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc
2281 ctgcccaggg accaccgacc caccaccggg aggtaagctg gctgcctcgc gcgtttcggt
2341 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
2401 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
2461 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
2521 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
2581 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
2641 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
2701 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
2761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
2821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
2881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
2941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
3001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
3061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
3121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
3181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
3241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
3301 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
3361 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
3421 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
3481 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
3541 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
3601 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
3661 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
3721 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
3781 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
3841 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
3901 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
3961 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
4021 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
4081 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
4141 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
4201 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
4261 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
4321 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
4381 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
4441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
4501 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
4561 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattgc
4621 tagcaattgc tagcaattgc tagcaattca taccagatca ccgaaaactg tcctccaaat
4681 gtgtccccct cacactccca aattcgcggg cttctgcctc ttagaccact ctaccctatt
4741 ccccacactc accggagcca aagccgcggc ccttccgttt ctttgctttt gaaagacccc
4801 acccgtaggt ggcaagctag cttaagtaac gccactttgc aaggcatgga aaaatacata
4861 actgagaata gaaaagttca gatcaaggtc aggaacaaag aaacagctga ataccaaaca
4921 ggatatctgt ggtaagcggt tcctgccccg gctcagggcc aagaacagat gagacagctg
4981 agtgatgggc caaacaggat atctgtggta agcagttcct gccccggctc ggggccaaga
5041 acagatggtc cccagatgcg gtccagccct cagcagtttc tagtgaatca tcagatgttt
5101 ccagggtgcc ccaaggacct gaaaatgacc ctgtacctta tttgaactaa ccaatcagtt
5161 cgcttctcgc ttctgttcgc gcgcttccgc tctccgagct caataaaaga gcccacaacc
5221 cctcactcgg cgcgccagtc ttccgataga ctgcgtcgcc cgggtacccg tattcccaat
5281 aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg ttccttggga gggtctcctc
5341 tgagtgattg actacccacg acgggggtct ttcatttggg ggctcgtccg ggatttggag
5401 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg
5461 tgtctgtccg attgtctagt gtctatgttt gatgttatgc gcctgcgtct gtactagtta
5521 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc
5581 cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga
5641 agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac
5701 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg
5761 tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg
5821 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac
5881 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
5941 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc
6001 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga
6061 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc
6121 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
6181 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
6241 ccttctctag gcgccggaat tcgcaccatg gtgagcaagg gcgaggagct gttcaccggg
6301 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
6361 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
6421 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
6481 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
6541 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
6601 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
6661 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
6721 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccac
//
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