| 质粒名称: | Cx43-sfGFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 70053 |
| 存储实验室: | David Spray |
|
载体骨架基本信息 |
|
| 载体名称: | EGFP-N1 |
| 空载体大小: | 5518 bp |
| 完整质粒大小: | 5893bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | JM109 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cx43 |
| 物种来源: | R. norvegicus (rat)褐家鼠(大鼠) |
| 插入大小: | 768 bp |
| 基因突变: | fused in frame to non-monomerized superfolder GFP |
| 启动子: | CMV |
| 标签/融合蛋白: | non-monomerized superfolder GFP (V207) (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CMV |
| 3’测序引物 | C1N1-rev: ACCTCTACAAATGTGGTATGGCTGATTATG |
|
文献引用方法 |
|
| 材料和方法部分: |
Cx43-sfGFP was a gift from David Spray (Addgene plasmid # 70053 ; http://n2t.net/addgene:70053 ; RRID:Addgene_70053)
|
| 参考文献部分: | Connexin Type and Fluorescent Protein-fusion Tag Determine Structural Stability of Gap Junction Plaques. Stout RF Jr, Snapp EL, Spray DC. J Biol Chem. 2015 Aug 11. pii: jbc.M115.659979. 10.1074/jbc.M115.659979 PubMed 26265468 |
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 Cx43-sfGFP 5893 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses rat Cx43 (Gja1 CDS) truncated at amino acid 258 and tagged
with non-monomerized sfGFP on the C-terminus. CMV promoter..
ACCESSION .
VERSION .
KEYWORDS Cx43-sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5893)
AUTHORS Stout RF Jr, Snapp EL, Spray DC
TITLE Connexin Type and Fluorescent Protein-fusion Tag Determine
Structural Stability of Gap Junction Plaques.
JOURNAL J Biol Chem. 2015 Aug 11. pii: jbc.M115.659979.
PUBMED 26265468
REFERENCE 2 (bases 1 to 5893)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5893
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 1833..1842
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1839..2558
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(1884..1905)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 2492..2513
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2681..2802
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2718..2737)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2772..2791
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2809..3264)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2946..2967)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3158..3177
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3291..3395
/gene="bla"
/label=AmpR promoter
promoter 3397..3754
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3420..3440)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3605..3740
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3667..3686
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3789..4583
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3843..3862)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4453..4472
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4771..4790)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4815..4862
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5191..5779
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5680..5699
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
661 gcaccatggg tgactggagt gccttgggga agcttctgga caaggtccaa gcctactcca
721 ccgctggagg gaaggtgtgg ctgtcagtgc tcttcatatt cagaatcctg ctcctgggga
781 cagctgttga gtcagcttgg ggtgatgaac agtctgcctt tcgctgtaac actcaacaac
841 ctggctgcga aaacgtctgc tatgacaagt ccttccccat ctctcacgtg cgcttctggg
901 tccttcagat catattcgtg tctgtgccca ccctcctgta cttggcccat gtgttctatg
961 tgatgaggaa ggaagagaag ctaaacaaga aagaagagga gctcaaagtg gcccagactg
1021 acggggtcaa cgtggagatg cacctgaagc agattgaaat caagaagttc aagtacggga
1081 ttgaagagca cggcaaggtg aaaatgaggg gcggcttgct gagaacctac atcatcagca
1141 tcctcttcaa gtctgtcttc gaggtggcct tcctgctcat ccagtggtac atctatgggt
1201 tcagcttgag cgcggtctac acctgcaaga gagatccctg cccccaccag gtagactgct
1261 tcctctcacg tcccacggag aaaaccatct tcatcatctt catgctggtg gtgtccttgg
1321 tgtctctcgc tttgaacatc attgagctct tctacgtctt cttcaaaggc gttaaggatc
1381 gcgtgaaggg aagaagcgat ccttaccacg ccaccactgg cccactgagc ccatcaaaag
1441 actgcggatc tccaaaatac gcctacttca atggctgctc ctcaccaacg gctccactct
1501 cgcctatgtc tcctcctggg tacaagctgg ttactggtga cagaaacaat tcctcgtgcc
1561 gcaattacaa caagcaagct agcgagcaaa actgggcgaa ctacagcgca gagcaaaatc
1621 gcatggggca ggccggaagc accatctcca actcgcacgc ccagccgttc gatttccccg
1681 acgacaacca gaatgccaaa aaagttgctg ctggacatga actccagcca ttagccatcg
1741 tggaccaacg accttccagc agagccagca gccgcgccag cagcaggcct cggcctgatg
1801 acctggagat ttccgcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
1861 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
1921 tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
1981 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
2041 gcgtgcagtg cttcagccgc taccccgacc acatgaagcg ccacgacttc ttcaagtccg
2101 ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca
2161 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
2221 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
2281 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga
2341 tccgccacaa cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
2401 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgtgc
2461 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
2521 ccgggatcac tcacggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
2581 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
2641 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
2701 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
2761 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
2821 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
2881 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
2941 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
3001 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
3061 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
3121 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
3181 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
3241 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
3301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
3361 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
3421 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
3481 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
3541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
3601 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
3661 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
3721 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
3781 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
3841 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
3901 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
3961 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
4021 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
4081 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
4141 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
4201 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
4261 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
4321 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
4381 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
4441 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
4501 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
4561 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
4621 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
4681 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
4741 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
4801 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
4861 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
4921 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
4981 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
5041 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
5101 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
5161 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
5221 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
5281 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
5341 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
5401 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
5461 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
5521 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
5581 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
5641 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
5701 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
5761 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
5821 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
5881 taccgccatg cat
//
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