| 质粒名称: | MCP-mCherry-SEC61B |
|---|---|
| 目录编号: | 207671 |
| 存储实验室: | Christine Mayr |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 完整质粒大小: | 6522 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Puromycin |
| 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: | SEC61B |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1350 bp |
| 启动子: | CMV |
| 标签/融合蛋白: | MCP-mCherry (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BmtI (未破坏) |
| 3’克隆位点: | NHindIII(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-SEC61B was a gift from Christine Mayr (Addgene plasmid # 207671 ; http://n2t.net/addgene:207671 ; RRID:Addgene_207671) |
| 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-SEC61B 6522 bp ds-DNA circular SYN 03-OCT-2023
DEFINITION To express fluorescently labeled MS2 coat binding protein (MCP)
fused with ER resident protein SEC61B.
ACCESSION .
VERSION .
KEYWORDS MCP-mCherry-SEC61B
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6522)
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 6522)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 3, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6522
/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..1962
/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"
CDS 1963..2250
/codon_start=1
/gene="SEC61B"
/product="human Sec61-beta subunit of the translocon in the
endoplasmic reticulum"
/label=Sec61-beta
/translation="PGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAGR
TTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
primer_bind complement(2362..2379)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2368..2592
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2638..3066
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2725..2744)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2935..2956
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3075..3095)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3080..3409
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3260..3395
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3322..3341
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3422..4021
/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(3422..3441)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3918..3938
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4195..4316
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4232..4251)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4286..4305
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4365..4381)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4365..4381)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4378..4400)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4389..4405
/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(4413..4443)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4458..4479
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4596..4613)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4767..5355)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4847..4866)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5526..6386)
/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 6149..6168
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6387..6491)
/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 agcctggtcc gacccccagt
1981 ggcactaacg tgggatcctc agggcgctct cccagcaaag cagtggccgc ccgggcggcg
2041 ggatccactg tccggcagag gaaaaatgcc agctgtggga caaggagtgc aggccgcaca
2101 acctcggcag gcaccggggg gatgtggcga ttctacacag aagattcacc tgggctcaaa
2161 gttggccctg ttccagtatt ggttatgagt cttctgttca tcgcttctgt atttatgttg
2221 cacatttggg gcaagtacac tcgttcgtag aagcttggta ccgagctcgg atccactagt
2281 ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc
2341 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
2401 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
2461 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
2521 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
2581 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
2641 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
2701 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
2761 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
2821 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
2881 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
2941 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
3001 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
3061 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
3121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
3181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
3241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
3301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
3361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccaac
3421 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
3481 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
3541 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
3601 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
3661 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
3721 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
3781 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
3841 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
3901 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
3961 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
4021 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
4081 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
4141 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
4201 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
4261 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
4321 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
4381 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
4441 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
4501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
4561 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
4621 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
4681 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
4741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
4801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
4861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
4921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
4981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
5041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
5101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
5161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
5221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
5281 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
5341 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
5401 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
5461 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
5521 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
5581 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
5641 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
5701 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
5761 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
5821 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
5881 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
5941 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
6001 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
6061 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
6121 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
6181 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
6241 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
6301 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
6361 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
6421 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
6481 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
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