| 质粒名称: | pTOM20-ddGFP-B |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 40291 |
| 存储实验室: | Robert Campbell |
|
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: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | TOMM20 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | ddGFP-B（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XhoI(未破坏) |
| 3' 克隆位点: | HindIII(未破坏) |
| 5' 测序引物 | TAATACGACTCACTATAGGG |
| 3' 测序引物 | TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pTOM20-ddGFP-B was a gift from Robert Campbell (Addgene plasmid # 40291 ; http://n2t.net/addgene:40291 ; RRID:Addgene_40291) |
| 参考文献部分: | Dimerization-dependent green and yellow fluorescent proteins. Alford SC, Ding Y, Simmen T, Campbell RE. ACS Synth Biol. 2012 Dec 21;1(12):569-75. doi: 10.1021/sb300050j. Epub 2012 Aug 14. 10.1021/sb300050j PubMed 23656278 |
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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 pTOM20-ddGFP-B 6523 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pTOM20-ddGFP-B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6523)
AUTHORS Alford SC, Ding Y, Simmen T, Campbell RE
TITLE Dimerization-dependent green and yellow fluorescent proteins.
JOURNAL ACS Synth Biol. 2012 Dec 21;1(12):569-75. doi: 10.1021/sb300050j.
Epub 2012 Aug 14.
PUBMED 23656278
REFERENCE 2 (bases 1 to 6523)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6523
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 167..546
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 547..750
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 701..721
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 795..814
/label=T7
/note="T7 promoter, forward primer"
promoter 795..813
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 838..847
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(1460..1478)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1728..1747
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 1922..1945
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(2046..2063)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2052..2276
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2322..2750
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2409..2428)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2619..2640
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2759..2779)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2764..3093
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2944..3079
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3006..3025
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3160..3954
/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(3214..3233)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3824..3843
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4128..4249
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4165..4184)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4219..4238
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4298..4314)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4298..4314)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4311..4333)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4322..4338
/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(4346..4376)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4391..4412
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4529..4546)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4700..5288)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4780..4799)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5459..6319)
/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 6082..6101
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6320..6424)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6499..6518)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
61 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
121 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
181 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
241 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
301 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
361 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
421 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
481 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
541 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
601 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
661 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
721 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
781 tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta gcgtgccgcc
841 accatggcct cgagaatggt gggtcggaac agcgccatcg ccgccggtgt atgcggggcc
901 cttttcattg ggtactgcat ctacttcgac cgcaaaagac gaagtgaccc caacttcaag
961 aacaggcttc gagaacgaag aaagaaacag aagcttgcca aggagagagc tgggctttcc
1021 aagttacctg accttaaaga tgctgaagct gttcagaagt tcttccttga agaaatacag
1081 cttggtgaag agttactagc tcaaggtgaa tatgagaagg gcgtagacca tctgacaaat
1141 gcaattgctg tgtgtggaca gccacagcag ttactgcagg tcttacagca aactcttcca
1201 ccaccagtgt tccagatgct tctgactaag ctcccaacaa ttagtcagag aattgtaagt
1261 gctcagagct tggctgaaga tgatgtggaa ggcggtagcg gggatccacc ggtcggtacc
1321 gcctcctccg aggacaacaa catggccacc atcaaagagt tcatgcgctt caaggtgcgc
1381 atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
1441 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gccctttgcc
1501 tgggacatcc tgtcccccca gatcatgtac ggctccaaag cgtacgtgag gcaccccgcc
1561 gacatccccg attacaagaa gctgcccttc cccgagggct tcaagtggga gcgcgtgatg
1621 aacttcgaag acggcggtct ggtgaccgtt acccaggact cctccctgca ggacggcacg
1681 ctgatctgca aggtgaagat tcgcggcacc aacttccccc ccgacggccc cgtaatgcag
1741 aagaagacca tgggctggga ggcctccacc gagatgctgt accccaagga cggcgtgctg
1801 aagggccata gctatcaggc cctgaagctg aaggacggcg gccactacct ggtggagttc
1861 gagaccatct acatggccaa gaagcccgtg caactgcccg gcgattactg tgtggacacc
1921 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgggcgctcc
1981 gagggccgcc accgtctggg catggacgag cggtacaagt aaaagcttgg taaaccgctg
2041 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
2101 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
2161 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
2221 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
2281 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
2341 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
2401 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
2461 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
2521 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
2581 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
2641 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
2701 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
2761 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
2821 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
2941 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
3001 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
3061 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
3121 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
3181 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
3241 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
3301 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
3361 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
3421 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
3481 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
3541 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
3601 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
3661 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
3721 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
3781 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
3841 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
3901 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
3961 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
4021 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
4081 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
4141 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
4201 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
4261 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
4321 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
4381 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
4441 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
4501 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
4561 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
4621 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
4681 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
4741 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
4801 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
4861 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctatagg tatctcagtt
4921 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
4981 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
5041 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
5101 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
5161 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
5221 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
5281 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
5341 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
5401 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
5461 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
5521 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
5581 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
5641 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
5701 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
5761 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
5821 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
5881 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
5941 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
6001 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
6061 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
6121 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
6181 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
6241 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
6301 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
6361 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
6421 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
6481 cctatggtgc actctcagta caatctgctc tgatgccgca tag
//
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