| 质粒名称: | mARG1 cassette 1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 134343 |
| 存储实验室: | Mikhail Shapiro |
|
载体骨架基本信息 |
|
| 载体名称: | pAF PB |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6756 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，合成生物学 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mARG 1 cassete 1,Mammalian acoustic reporter gene1 cassette 1 |
| 物种来源: | Bacillus megaterium |
| 插入大小: | 1571 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | TRE3G |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | AAAGGAAAACCACGTCCCCGT |
| 3' 测序引物 | CCCAGAAGGTACCCCATTGTA |
|
文献引用方法 |
|
| 材料和方法部分: | mARG1 cassette 1 was a gift from Mikhail Shapiro (Addgene plasmid # 134343 ; http://n2t.net/addgene:134343 ; RRID:Addgene_134343) |
| 参考文献部分: | Ultrasound imaging of gene expression in mammalian cells. Farhadi A, Ho GH, Sawyer DP, Bourdeau RW, Shapiro MG. Science. 2019 Sep 27;365(6460):1469-1475. doi: 10.1126/science.aax4804. 10.1126/science.aax4804 PubMed 31604277 |
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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 mARG1 cassette 1 6756 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian acoustic reporter gene 1 cassette 1 on the piggyBac
trasposon plasmid..
ACCESSION .
VERSION .
KEYWORDS mARG1 cassette 1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6756)
AUTHORS Farhadi A, Ho GH, Sawyer DP, Bourdeau RW, Shapiro MG
TITLE Ultrasound imaging of gene expression in mammalian cells.
JOURNAL Science. 2019 Sep 27;365(6460):1469-1475. doi:
10.1126/science.aax4804.
PUBMED 31604277
REFERENCE 2 (bases 1 to 6756)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6756
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 78..140
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
promoter 574..923
/label=TRE3GV promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, optimized for
retroviral and lentiviral vectors"
protein_bind 582..600
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 618..636
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 654..672
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 690..708
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 726..744
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 762..780
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 798..816
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 862..886
/label=LNCX
/note="Human CMV promoter, forward primer"
misc_feature 1240..1791
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1384..1401)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 1611..1630
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 1803..2513
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(1948..1966)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2216..2235
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2530..2651
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2567..2586)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2621..2640
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
repeat_region 3414..3448
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind complement(3939..3955)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3939..3955)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3952..3974)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3963..3979
/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(3987..4017)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4032..4053
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4170..4187)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4341..4929)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4421..4440)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5100..5960)
/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 5723..5742
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5961..6065)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6091..6546)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6228..6249)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6440..6459
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6673..6695
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6687..6704
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6688..6704
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 gcggtgcaaa tgtgttttac agcgtgatgg agcagatgaa gatgctcgac acgctgcaga
61 acacgcagct agattaaccc tagaaagata atcatattgt gacgtacgtt aaagataatc
121 atgtgtaaaa ttgacgcatg tgttttatcg gtctgtatat cgaggtttat ttattaattt
181 gaatagatat taagttttat tatatttaca cttacatact aataataaat tcaacaaaca
241 atttatttat gtttatttat ttattaaaaa aaacaaaaac tcaaaatttc ttctataaag
301 taacaaaact tttatgaggg acagcccccc cccaaagccc ccagggatgt aattacgtcc
361 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
421 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
481 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
541 gggaaaaggc ctccacggcc actagtttca ctcgagttta ctccctatca gtgatagaga
601 acgtatgaag agtttactcc ctatcagtga tagagaacgt atgcagactt tactccctat
661 cagtgataga gaacgtataa ggagtttact ccctatcagt gatagagaac gtatgaccag
721 tttactccct atcagtgata gagaacgtat ctacagttta ctccctatca gtgatagaga
781 acgtatatcc agtttactcc ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac
841 ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct ggagcaattc
901 cacaacactt ttgtcttata cttggtacct atgcatgcca ccatgagcat ccagaagtcc
961 accaacagca gcagcctggc cgaagtgatc gaccggatcc tggacaaggg catcgtgatc
1021 gacgccttcg ccagagtgtc cgtcgtgggc atcgagatcc tgaccatcga ggccagagtc
1081 gtgatcgcca gcgtggacac ctggctgaga tatgccgaag ccgtgggcct gctgcgggac
1141 gacgtggaag aaaatggcct gcccgagcgg agcaacagct ctgagggaca gccccggttc
1201 agcatctgaa ctaaatcgca ctgtcggcgt ccccccctaa cgttactggc cgaagccgct
1261 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
1321 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
1381 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
1441 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
1501 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
1561 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
1621 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
1681 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg
1741 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
1801 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
1861 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
1921 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc
1981 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
2041 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
2101 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
2161 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
2221 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
2281 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
2341 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
2401 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
2461 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tgaactagtt
2521 cgttaactaa acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
2581 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
2641 aatgtatctt atcatgtctg gaattgactc aaatgatgtc aattagtcta tcagaagctc
2701 atctggtctc ccttccgggg gacaagacat ccctgtttaa tatttaaaca gcagtgttcc
2761 caaactgggt tcttatatcc cttgctctgg tcaaccaggt tgcagggttt cctgtcctca
2821 caggaacgaa gtccctaaag aaacagtggc agccaggttt agccccggaa ttgactggat
2881 tcctttttta gggcccattg gtatggcttt ttccccgtat ccccccaggt gtctgcaggc
2941 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
3001 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
3061 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
3121 ttgggggggg gctgtccctg atatctataa caagaaaata tatatataat aagttatcac
3181 gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt
3241 aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac
3301 ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat
3361 gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt
3421 caattttacg cagactatct ttctagggtt aatctagctg catcaggatc atatcgtcgg
3481 gtcttttttc cggctcagtc atcgcccaag ctggcgctat ctgggcatcg gggaggaaga
3541 agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg aggatgactg
3601 ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg atgattcggg aaggtgtggc
3661 catgcacgcc tttaacggtg aactgttcgt tcaggccacc tgggatacca gttcgtcgcg
3721 gcttttccgg acacagttcc ggatggtcag cccgaagcgc atcagcaacc cgaacaatac
3781 cggcgacagc cggaactgcc gtgccggtgt gcagattaat gacagcggtg cggcgctggg
3841 atattacgtc agcgaggacg ggtatcctgg ctggatgccg cagaaatgga catggatacc
3901 ccgtgagtta cccggcgggc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
3961 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
4021 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
4081 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
4141 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
4201 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
4261 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4321 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4381 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4441 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
4501 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
4561 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4621 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
4681 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
4741 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
4801 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
4861 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
4921 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
4981 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
5041 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
5101 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
5161 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
5221 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
5281 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
5341 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
5401 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
5461 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
5521 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
5581 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
5641 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
5701 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
5761 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
5821 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
5881 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
5941 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
6001 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
6061 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
6121 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
6181 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
6241 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
6301 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
6361 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
6421 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
6481 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
6541 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
6601 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
6661 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcctcgttca
6721 ttcacgtttt tgaacccgtg gaggacgggc agactc
//
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