| 质粒名称: | 2C::tdTomato Reporter |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 28027 |
| 存储实验室: | Samuel Pfaff |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA Hygro |
| 载体出品公司: | Invitrogen/Life Tech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6400 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 潮霉素 |
| 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: | 2C Promoter (2-cell stage promoter) |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 730 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | tdTomato |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Mlu1(未破坏) |
| 3' 克隆位点: | HindIII (未破坏) |
| 5' 测序引物 | Amp-R |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | 2C::tdTomato Reporter was a gift from Samuel Pfaff (Addgene plasmid # 40281 ; http://n2t.net/addgene:40281 ; RRID:Addgene_40281) |
| 参考文献部分: | Embryonic stem cell potency fluctuates with endogenous retrovirus activity. Macfarlan TS, Gifford WD, Driscoll S, Lettieri K, Rowe HM, Bonanomi D, Firth A, Singer O, Trono D, Pfaff SL. Nature. 2012 Jul 5;487(7405):57-63. 10.1038/nature11244 PubMed 22722858 |
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载体基本信息
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载体简介
载体序列
LOCUS 2C::tdTomato Reporter 7132 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS 2C::tdTomato Reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7132)
AUTHORS Macfarlan TS, Gifford WD, Driscoll S, Lettieri K, Rowe HM, Bonanomi
D, Firth A, Singer O, Trono D, Pfaff SL
TITLE Embryonic stem cell potency fluctuates with endogenous retrovirus
activity.
JOURNAL Nature. 2012 Jul 5;487(7405):57-63.
PUBMED 22722858
REFERENCE 2 (bases 1 to 7132)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7132
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(75..94)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS 1067..2497
/codon_start=1
/product="tandem dimeric (pseudo-monomeric) derivative of
DsRed (Shaner et al., 2004)"
/label=tdTomato
/note="mammalian codon-optimized"
/translation="MVSKGEDVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
RSEGRHHLFLYGMDELYK"
primer_bind complement(2585..2602)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2591..2815
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2861..3289
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2948..2967)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3158..3179
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3298..3318)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3303..3632
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3483..3618
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3545..3564
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3681..4706
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
polyA_signal 4836..4957
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4873..4892)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4927..4946
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5006..5022)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5006..5022)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5019..5041)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5030..5046
/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(5054..5084)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5099..5120
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5237..5254)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5408..5996)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5488..5507)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6167..7027)
/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 6790..6809
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7028..7132)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
61 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
121 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
181 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
241 gatgtacggg ccagatatac gcgtgctagc tgtagtggtt attcctggtt gtcaacttga
301 caatatttgg aatgaactac agtccggaat tggaaggctc accagtgacc cttatctgga
361 ggcttggaga tccttatctg gatcttggtt tgaagatctt gagccatagt ggctatggat
421 tccagaagat tgaatctccg agtttaagga acacaccttt aatctgggct acgcctttca
481 tctgggatta aaggtgtggt ggaacacacc tttaatctgg gctacacctt ctgctggaga
541 caatataagg acattggaag aagggagtct agctcttgct cctgctcctt cgcctgcttg
601 ctgcgtgaga ctgagtaact gctagatcct tggacttcca ttcacagctg tgactgaacg
661 attgttggga attgagctgc cgactgtaag tcatcaataa attcctttat tatctagaga
721 ctatccataa gttctgtgac tctagagaac cctgactaat acagaagttg gtaccaggag
781 tggttctaga gtaacagaag tacaaggatg aatcttttaa aattctggaa ttggcttgtt
841 gatccaccag cactttcaac tattgaaacc tctccagatt ctctccctcc tgggagctca
901 gagaattttg aagacccatg gttgaaacta tattccgaac ttaaagaagc taatgccctt
961 gattttctta atgaattagg tgattcagtg cacaaagctt ggtaccgagc tcggatccac
1021 tagtaacggc cgccagtgtg ctggaattcg cccttgtctg tctagaatgg tgagcaaggg
1081 cgaggacgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
1141 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
1201 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
1261 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
1321 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
1381 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
1441 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
1501 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
1561 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
1621 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
1681 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
1741 cctggggcat ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga
1801 caacaacatg gccgtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat
1861 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
1921 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
1981 cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta
2041 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
2101 cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt
2161 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
2221 ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
2281 ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat
2341 ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac
2401 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
2461 cctgttcctg tacggcatgg acgagctgta caagtaatct agacccccaa gggcgaattc
2521 tgcagatatc catcacactg gcggccgctc gagtctagag ggcccgttta aacccgctga
2581 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
2641 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
2701 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
2761 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
2821 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
2881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
2941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
3001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
3061 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
3121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
3181 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
3241 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
3301 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
3361 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
3421 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
3481 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
3541 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
3601 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
3661 tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
3721 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
3781 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
3841 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
3901 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
3961 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
4021 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
4081 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
4141 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
4201 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
4261 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
4321 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
4381 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
4441 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
4501 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
4561 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
4621 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
4681 gcactcgtcc gagggcaaag gaatagcacg tgctacgaga tttcgattcc accgccgcct
4741 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
4801 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
4861 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
4921 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
4981 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
5041 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
5101 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
5161 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
5221 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
5281 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
5341 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
5401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
5461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
5521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
5581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
5641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
5701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
5761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
5821 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
5881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
5941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
6001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
6061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
6121 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
6181 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
6241 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
6301 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
6361 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
6421 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
6481 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
6541 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
6601 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
6661 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
6721 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
6781 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
6841 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
6901 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
6961 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
7021 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
7081 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cg
//
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