| 質粒名稱: | Str-KDEL_ST-SBP-EGFP |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 65264 |
| 存儲實驗室: | Franck Perez |
|
載體骨架基本信息 |
|
| 載體名稱: | pIRESneo3 |
| 載體出品公司: | Clontech |
| 完整質粒大小: | 6044bp |
| 載體類型: | 哺乳細胞表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | Str-KDEL and truncated sialyltransferas、ST6GAL1 |
| 物種來源: | H. sapiens (human) |
| 基因突變: |
contains only amino acids 1 to 43 of ST
Entrez GeneST6GAL1 (a.k.a. CDw75, SIAT1, ST6GalI, ST6N)
|
| GenBank編號 | JN247409 |
| 啟動子: | CMV |
| 標簽/融合蛋白: |
Streptavidin Binding Protein (SBP) (C terminal on insert)
EGFP (C terminal on insert)
|
| 克隆方法: | 未知 |
| 5’克隆位點: | IRES-F |
| 3’克隆位點: | EGFP-N |
|
文獻引用方法 |
|
| 材料和方法部分: | Str-KDEL_ST-SBP-EGFP was a gift from Franck Perez (Addgene plasmid # 65264 ; //n2t.net/addgene:65264 ; RRID:Addgene_65264) |
| 參考文獻部分: | Synchronization of secretory protein traffic in populations of cells. Boncompain G, Divoux S, Gareil N, de Forges H, Lescure A, Latreche L, Mercanti V, Jollivet F, Raposo G, Perez F. Nat Methods. 2012 Mar 11;9(5):493-8. doi: 10.1038/nmeth.1928. 10.1038/nmeth.1928 PubMed 22406856 |
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Str-KDEL_ST-SBP-EGFP載體質粒基本信息
Str-KDEL_ST-SBP-EGFP質粒圖譜載體圖譜和Str-KDEL_ST-SBP-EGFP載體序列質粒序列多克隆位點信息
Str-KDEL_ST-SBP-EGFP質粒載體簡介
Str-KDEL_ST-SBP-EGFP質粒序列載體序列
LOCUS Str-KDEL_ST-SBP-EGFP 6044 bp DNA circular SYN 14-APR-2025
DEFINITION synchronize trafficking of ST from the ER (RUSH system).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6044)
AUTHORS Boncompain G, Divoux S, Gareil N, de Forges H, Lescure A, Latreche
L, Mercanti V, Jollivet F, Raposo G, Perez F
TITLE Synchronization of secretory protein traffic in populations of
cells.
JOURNAL Nat Methods. 2012 Mar 11;9(5):493-8. doi: 10.1038/nmeth.1928.
PUBMED 22406856
REFERENCE 2 (bases 1 to 6044)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nmeth";
journalName: "Nat Methods"; date: "2012-03-11- 11"; volume: "9";
issue: "5"; pages: "493-8"
COMMENT Sequence Label: Str-KDEL_ST-SBP-EGFP
FEATURES Location/Qualifiers
source 1..6044
/mol_type="other DNA"
/organism="synthetic DNA construct"
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"
intron 1646..1875
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
primer_bind 1858..1877
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature join(1933..2507,2508..2510,2511..2519)
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2100..2117)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2327..2346
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2671..2784
/codon_start=1
/product="streptavidin-binding peptide"
/label=SBP
/note="selected from a peptide library; binds streptavidin
with nanomolar affinity (Keefe et al., 2001)"
/translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
CDS join(2794..2796,2797..2799,2800..3510)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(2794..2796,2797..2799,2800..3510)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2839..2860)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3100..3119)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3447..3468
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 3664..3785
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3701..3720)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3887..3903)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3887..3903)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3900..3922)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3911..3927
/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(join(3935..3941,3942..3959,3960..3965))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3980..4001
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4118..4135)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4289..4877)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4369..4388)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5048..5839,5840..5908))
/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 5671..5690
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5909..6013)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgacc
1081 ctagcaaaga ctcaaaagct caggtgtccg ctgccgaggc tggcattact ggaacatggt
1141 acaatcagct cgggagcacc tttattgtga ctgctggagc cgatggagcc ctcaccggaa
1201 catacgaatc tgctgtggga aacgccgaat cacggtacgt cctcactggc cgatacgata
1261 gtgcccctgc caccgacgga tctgggactg ccctgggatg gactgtcgct tggaaaaaca
1321 actaccggaa tgctcattct gccacaacat ggagtggaca gtacgtggga ggcgctgagg
1381 ctagaatcaa tacacagtgg ctgctcacat ctggcacaac cgaggcaaat gcttggaaat
1441 ccaccctggt gggacatgac acattcacca aagtgaaacc ctccgccgct tcaatcgatg
1501 ccgccaaaaa agccggagtc aacaacggca atcctctgga tgccgtccag cagctcggac
1561 cgaaagatga actgtgagcg gccgcataga taactgatcc agtgtgctgg aattaattcg
1621 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
1681 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
1741 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
1801 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
1861 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
1921 cggccaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
1981 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
2041 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
2101 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
2161 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
2221 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
2281 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
2341 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
2401 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
2461 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
2521 gggaggcgcg ccatgattca caccaacctg aagaaaaagt tcagctgctg cgtcctggtc
2581 tttcttctgt ttgcagtcat ctgtgtgtgg aaggaaaaga agaaagggag ttactatgat
2641 tcctttaaat tgcaaaccaa ggggaattcc atggacgaga agaccactgg ttggcgaggt
2701 ggacacgttg ttgaaggact ggctggggaa cttgaacaac ttcgtgcacg actggagcat
2761 cacccacaag gtcaacgtga accacctgca ggtatggtga gcaaaggcga ggagctgttc
2821 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
2881 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
2941 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
3001 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
3061 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
3121 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
3181 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
3241 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
3301 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
3361 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
3421 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
3481 atcactctcg gcatggacga gctgtacaag ggccggcctt aaggcctcga gggccaatta
3541 attaactcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
3601 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
3661 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
3721 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
3781 tcttaacgcg tcgagtgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
3841 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
3901 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
3961 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
4021 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
4081 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
4141 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
4201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
4261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
4321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
4381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
4441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
4501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
4561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
4621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
4681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
4741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
4801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
4861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
4921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
4981 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
5041 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
5101 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
5161 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
5221 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
5281 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
5341 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
5401 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
5461 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
5521 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
5581 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
5641 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
5701 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
5761 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
5821 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
5881 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
5941 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
6001 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
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