| 质粒名称: | pLV-EF1a-MinE-IRES-Blast |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 213115 |
| 存储实验室: | Scott Coyle |
|
载体骨架基本信息 |
|
| 载体名称: | pLV-EF1a |
| 载体出品公司: | Clontech |
| 空载体大小: | 8528bp |
| 完整质粒大小: | 8795bp |
| 载体类型: | 哺乳细胞表达, Lentiviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | MinE |
| 插入大小: | 267 bp |
| 启动子: | EF1a |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | EF1a_F (TCAAGCCTCAGACAGTGGTTC) |
| 3’测序引物 | EF1a_R (TAGAGGGAAACCGTTGCTAGC) |
|
文献引用方法 |
|
| 材料和方法部分: |
pLV-EF1a-MinE-IRES-Blast was a gift from Scott Coyle (Addgene plasmid # 213115 ; http://n2t.net/addgene:213115 ; RRID:Addgene_213115)
|
| 参考文献部分: | A programmable reaction-diffusion system for spatiotemporal cell signaling circuit design. Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM. Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi: 10.1016/j.cell.2023.12.007. 10.1016/j.cell.2023.12.007 PubMed 38181787 |
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 pLV-EF1a-MinE-IRES-Blast 8795 bp ds-DNA circular SYN 26-JAN-2024
DEFINITION Expression of untagged MinE in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLV-EF1a-MinE-IRES-Blast
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8795)
AUTHORS Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM
TITLE A programmable reaction-diffusion system for spatiotemporal cell
signaling circuit design.
JOURNAL Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi:
10.1016/j.cell.2023.12.007.
PUBMED 38181787
REFERENCE 2 (bases 1 to 8795)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 26, 2024 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8795
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(202..213)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 526..1159
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(1287..1303)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1287..1303)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1300..1322)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1311..1327
/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(1335..1365)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1380..1401
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1518..1535)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1689..2277)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1769..1788)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2448..3308)
/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 3071..3090
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3309..3413)
/gene="bla"
/label=AmpR promoter
polyA_signal 3461..3595
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3498..3517)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3552..3571
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 3763..4396
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 4443..4568
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5065..5298
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 5483..5527
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 5790..5905
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 5983..6194
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
intron 6195..7133
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 7090..7110
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 7149..7158
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(7666..7683)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 7893..7912
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
misc_feature 8526..319
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8579..8599)
/label=WPRE-R
/note="WPRE, reverse primer"
ORIGIN
1 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
61 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
121 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
181 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
241 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
301 ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag
361 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
421 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
481 aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc
541 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
601 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt
661 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca
721 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag
781 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt
841 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag
901 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag
961 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
1021 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
1081 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
1141 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
1201 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt accgtcgacc
1261 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
1321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
1381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
1441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
1501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
1561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
1621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
1681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
1741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
1801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
1861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
1921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
1981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
2041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
2101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
2161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
2221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
2281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
2341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
2401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
2461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
2521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
2581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
2641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
2701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
2761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
2821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
2881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
2941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
3001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
3061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
3121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
3181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
3241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
3301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
3361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
3421 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
3481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
3541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
3601 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
3661 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
3721 aagaaattgt atttgttaaa tatgtactac aaacttagta gttggaaggg ctaattcact
3781 cccaaagaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
3841 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
3901 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
3961 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
4021 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
4081 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
4141 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
4201 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
4261 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
4321 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
4381 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
4441 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
4501 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
4561 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
4621 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
4681 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
4741 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
4801 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
4861 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
4921 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
4981 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
5041 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
5101 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
5161 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
5221 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
5281 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
5341 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
5401 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
5461 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
5521 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
5581 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
5641 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
5701 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
5761 attagtgaac ggatctcgac ggtatcgcct ttaaaagaaa aggggggatt ggggggtaca
5821 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
5881 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
5941 atcgatgagg ccctttcgtc ttcactcgag gtgcccgtca gtgggcagag cgcacatcgc
6001 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
6061 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
6121 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
6181 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
6241 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
6301 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
6361 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
6421 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
6481 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
6541 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
6601 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
6661 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
6721 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
6781 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
6841 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
6901 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
6961 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
7021 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
7081 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
7141 gaggatccgc caccatggcc cttcttgact tcttcctgtc tcgaaaaaag aacacagcga
7201 atatcgcgaa agaacggttg caaattatag tggcggagag acgacgaagc gacgcggagc
7261 ctcattacct tcctcaattg cgaaaagaca tactggaagt gatctgcaag tacgttcaaa
7321 ttgaccctga gatggtgacc gtgcaacttg aacagaagga tggagatata agcatcctgg
7381 aattgaacgt aacccttccc gaggcggaag aacttaaata agcggccgct ctagagtcga
7441 cgggccgcgg taacaattgt taactaactt aagctagcaa cggtttccct ctagcgggat
7501 caattccgcc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
7561 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
7621 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
7681 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
7741 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
7801 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
7861 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
7921 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
7981 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
8041 acgtggtttt cctttgaaaa acacgataat accatggtca tgaaaacatt taacatttct
8101 caacaagatc tagaattagt agaagtagcg acagagaaga ttacaatgct ttatgaggat
8161 aataaacatc atgtgggagc ggcaattcgt acgaaaacag gagaaatcat ttcggcagta
8221 catattgaag cgtatatagg acgagtaact gtttgtgcag aagccattgc gattggtagt
8281 gcagtttcga atggacaaaa ggattttgac acgattgtag ctgttagaca cccttattct
8341 gacgaagtag atagaagtat tcgagtggta agtccttgtg gtatgtgtag ggagttgatt
8401 tcagactatg caccagattg ttttgtgtta atagaaatga atggcaagtt agtcaaaact
8461 acgattgaag aactcattcc actcaaatat acccgaaatt aacccgggcg gggcgcgtct
8521 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
8581 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
8641 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
8701 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
8761 ccactggttg gggcattgcc accacctgtc agctc
//
