| 质粒名称: | MCP-mCherry |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 207668 |
| 存储实验室: | Christine Mayr |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Puromycin |
| 完整质粒大小: | 6163 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | MCP-mCherry |
| 物种来源: | Synthetic |
| 插入大小: | 1065 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BmtI(未破坏) |
| 3’克隆位点: | BsrGI(未破坏) |
| 5’测序引物 | T7 |
| 3’测序引物 | BGHR |
|
文献引用方法 |
|
| 材料和方法部分: | MCP-mCherry was a gift from Christine Mayr (Addgene plasmid # 207668 ; http://n2t.net/addgene:207668 ; RRID:Addgene_207668) |
| 参考文献部分: | Subcytoplasmic location of translation controls protein output. Horste EL, Fansler MM, Cai T, Chen X, Mitschka S, Zhen G, Lee FCY, Ule J, Mayr C. bioRxiv 2022.11.04.515216 10.1101/2022.11.04.515216 |
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 MCP-mCherry 6163 bp ds-DNA circular SYN 27-SEP-2023
DEFINITION To express fluorescently labeled MS2 coat binding protein (MCP).
ACCESSION .
VERSION .
KEYWORDS MCP-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6163)
AUTHORS Horste EL, Fansler MM, Cai T, Chen X, Mitschka S, Zhen G, Lee FCY,
Ule J, Mayr C
TITLE Subcytoplasmic location of translation controls protein output
JOURNAL bioRxiv 2022.11.04.515216
REFERENCE 2 (bases 1 to 6163)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6163
/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 1258..1965
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(1400..1418)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1668..1687
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(2003..2020)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2009..2233
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2279..2707
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2366..2385)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2576..2597
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2716..2736)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2721..3050
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2901..3036
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2963..2982
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3063..3662
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3063..3082)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3559..3579
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 3836..3957
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3873..3892)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3927..3946
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4006..4022)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4006..4022)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4019..4041)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4030..4046
/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(4054..4084)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4099..4120
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4237..4254)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4408..4996)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4488..4507)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5167..6027)
/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 5790..5809
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6028..6132)
/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 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
961 gtcgccccaa gcaacttcgc taacgggatc gctgaatgga tcagctctaa ctcgcgttca
1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
1081 atcaaagtcg aggtgcctaa aggcgcctgg cgttcgtact taaatatgga actaaccatt
1141 ccaattttcg ccacgaattc cgactgcgag cttattgtta aggcaatgca aggtctccta
1201 aaagatggaa acccgattcc ctcagcaatc gcagcaaact ccggcatcta cggatccgtg
1261 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
1321 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
1381 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
1441 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
1501 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
1561 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
1621 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
1681 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
1741 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
1801 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
1861 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
1921 gagggccgcc actccaccgg cggcatggac gagctgtaca agtagctcga gtctagaggg
1981 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
2041 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
2101 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
2161 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
2221 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
2281 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
2341 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
2401 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
2461 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
2521 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
2581 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
2641 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
2701 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
2821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
2881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
2941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
3001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccaa
3061 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
3121 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
3181 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
3241 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
3301 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
3361 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
3421 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
3481 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
3541 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
3601 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
3661 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
3721 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
3781 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
3841 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
3901 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
3961 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
4021 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
4081 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
4141 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
4201 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
4261 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
4321 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
4381 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
4441 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
4501 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
4561 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
4621 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
4681 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
4741 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
4801 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
4861 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
4921 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
4981 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
5041 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
5101 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
5161 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
5221 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
5281 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
5341 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
5401 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
5461 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
5521 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
5581 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
5641 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
5701 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
5761 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
5821 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
5881 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
5941 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
6001 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
6061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
6121 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
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