| 质粒名称: | Laconic/pcDNA3.1(-) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 44238 |
| 存储实验室: | Luis Felipe Barros |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1(-) |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5418bp |
| 完整质粒大小: | 7648bp |
| 载体修饰: | 哺乳动物表达；乳酸生物传感器 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | Laconic (LACtate Optical Nano Indicator from CECs) |
| 物种来源: | 合成的 |
| 插入大小: | 2230 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| Laconic/pcDNA3.1(-) was a gift from Luis Felipe Barros (Addgene plasmid # 44238 ; http://n2t.net/addgene:44238 ; RRID:Addgene_44238) | |
| A Genetically Encoded FRET Lactate Sensor and Its Use To Detect the Warburg Effect in Single Cancer Cells. San_Martin A, Ceballo S, Ruminot I, Lerchundi R, Frommer WB, Barros LF. PLoS One. 2013;8(2):e57712. doi: 10.1371/journal.pone.0057712. Epub 2013 Feb 26. 10.1371/journal.pone.0057712 PubMed 23469056 | |
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
质粒图谱.png)
载体简介
载体序列
LOCUS Laconic/pcDNA3.1(-) 7651 bp ds-DNA circular SYN 30-JUL-2019
DEFINITION a genetically-encoded sensor for lactate concentration, transport
and metabolic flux.
ACCESSION .
VERSION .
KEYWORDS Laconic/pcDNA3.1(-)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7651)
AUTHORS San_Martin A, Ceballo S, Ruminot I, Lerchundi R, Frommer WB, Barros
LF
TITLE A Genetically Encoded FRET Lactate Sensor and Its Use To Detect the
Warburg Effect in Single Cancer Cells.
JOURNAL PLoS One. 2013;8(2):e57712. doi: 10.1371/journal.pone.0057712. Epub
2013 Feb 26.
PUBMED 23469056
REFERENCE 2 (bases 1 to 7651)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 30, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7651
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 987..1694
/codon_start=1
/product="monomeric teal (cyan) fluorescent protein (Ai et
al., 2006)"
/label=mTFP1
/note="mammalian codon-optimized"
/translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
VARNSTDGMDELYK"
CDS 2496..3215
/codon_start=1
/product="Venus YFP with monomerizing A206K mutation (Nagai
et al., 2002)"
/label=mVenus
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2541..2562)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2802..2821)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3149..3170
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3242..3259)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3248..3472
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3518..3946
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3605..3624)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3815..3836
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3955..3975)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3960..4289
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4140..4275
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4202..4221
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4356..5150
/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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4410..4429)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5020..5039
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5324..5445
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5361..5380)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5415..5434
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5494..5510)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5494..5510)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5507..5529)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5518..5534
/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(5542..5572)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5587..5608
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5725..5742)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5896..6484)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5976..5995)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6655..7515)
/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 7278..7297
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7516..7620)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc
961 accacactgg actagtggat ccgaacatgg tgagcaaggg cgaggagacc acaatgggcg
1021 taatcaagcc cgacatgaag atcaagctga agatggaggg caacgtgaat ggccacgcct
1081 tcgtgatcga gggcgagggc gagggcaagc cctacgacgg caccaacacc atcaacctgg
1141 aggtgaagga gggagccccc ctgcccttct cctacgacat tctgaccacc gcgttcgcct
1201 acggcaacag ggccttcacc aagtaccccg acgacatccc caactacttc aagcagtcct
1261 tccccgaggg ctactcttgg gagcgcacca tgaccttcga ggacaagggc atcgtgaagg
1321 tgaagtccga catctccatg gaggaggact ccttcatcta cgagatacac ctcaagggcg
1381 agaacttccc ccccaacggc cccgtgatgc agaagaagac caccggctgg gacgcctcca
1441 ccgagaggat gtacgtgcgc gacggcgtgc tgaagggcga cgtcaagcac aagctgctgc
1501 tggagggcgg cggccaccac cgcgttgact tcaagaccat ctacagggcc aagaaggcgg
1561 tgaagctgcc cgactatcac tttgtggacc accgcatcga gatcctgaac cacgacaagg
1621 actacaacaa ggtgaccgtt tacgagagcg ccgtggcccg caactccacc gacggcatgg
1681 acgagctgta caagagatct ggtaccatga ttgttttacc cagacgcctg tcagacgagg
1741 ttgccgatcg tgtgcgggcg ctgattgatg aaaaaaacct ggaagcgggc atgaagttgc
1801 ccgctgagcg ccaactggcg atgcaactcg gcgtatcacg taattcactg cgcgaggcgc
1861 tggcaaaact ggtgagtgaa ggcgtgctgc tcagtcgacg cggcggcggg acgtttattc
1921 gctggcgtca tgacacatgg tcggagcaaa acatcgtcca gccgctaaaa acactgatgg
1981 ccgatgatcc ggattacagt ttcgatattc tggaagcccg ctacgccatt gaagccagca
2041 ccgcatggca tgcggcaatg cgcgccacac ctggcgacaa agaaaagatt cagctttgct
2101 ttgaagcaac gctaagtgaa gacccggata tcgcctcaca agcggacgtt cgttttcatc
2161 tggcgattgc cgaagcctca cataacatcg tgctgctgca aaccatgcgc ggtttcttcg
2221 atgtcctgca atcctcagtg aagcatagcc gtcagcggat gtatctggtg ccaccggttt
2281 tttcacaact gaccgaacaa catcaggctg tcattgacgc catttttgcc ggtgatgctg
2341 acggggcgcg taaagcaatg atggcgcacc ttagttttgt tcacaccacc atgaaacgat
2401 tcgatgaaga tcaggctcgc cacgcacgga ttacccgcct gcccggtgag cataatgagc
2461 attcgaggga gaaaaacgca cttaagagat ctaccatggt gagcaagggc gaggagctgt
2521 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
2581 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
2641 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc
2701 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
2761 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
2821 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
2881 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
2941 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
3001 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
3061 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
3121 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
3181 ggatcactct cggcatggac gagctgtaca agtaaaagct taagtttaaa ccgctgatca
3241 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
3301 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
3361 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
3421 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
3481 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
3541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
3601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
3661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
3721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
3781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
3841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
3901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
4201 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
4261 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
4321 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
4381 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
4441 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
4501 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
4561 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
4621 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
4681 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
4741 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
4801 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
4861 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
4921 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
4981 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
5041 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
5101 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
5161 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
5221 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
5281 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
5341 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
5401 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
5461 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
5521 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
5581 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
5641 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
5701 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
5761 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
5821 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
5881 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
5941 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
6001 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
6061 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
6121 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
6181 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
6241 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
6301 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
6361 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
6421 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
6481 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
6541 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
6601 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
6661 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
6721 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
6781 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
6841 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
6901 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
6961 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
7021 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
7081 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
7141 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
7201 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
7261 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
7321 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
7381 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
7441 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
7501 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
7561 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
7621 cacatttccc cgaaaagtgc cacctgacgt c
//
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