- 商品货号:FH1307
- 规格:10μl质粒
- 启动子:ECMV promoter
- 复制子:pUC ori,SV40 ori
- 终止子:WPRE
- 质粒分类:病毒系列质粒;慢病毒类质粒;病毒表达质粒
- 质粒大小:8538bp
- 原核抗性:Amp(100μg/ml)
- 筛选标记:ZsGreen1,Puro
- 克隆菌株:Stbl3
- 表达宿主:293T等哺乳细胞
- 诱导方式:无需诱导
- 5测序引物:CMV-F(CGCAAATGGGCGGTAGGCGTG)
- 3测序引物:EF1-R(CTAGGCACCCGTTCAATTGC)
- 备注:本质粒由武汉淼灵生物科技有限公司分享至淼灵质粒平台。
- 质粒简介: pCDH-ECMV-MCS-3×FLAG-EF1α-ZsGreen1-T2A-Puro质粒是在pCDH载体的基础上改造而成,CMV启动目的基因表达,C端的3×FLAG融合标签能很好的被FLAG抗体识别。该质粒可作为过表达质粒直接转染哺乳细胞,也可包装成慢病毒侵染细胞。质粒本身带有ZsGreen1荧光蛋白和Puro筛选标记,可利用荧光显微镜观察转染效率,也可用嘌呤霉素筛选转染后的细胞得到阳性克隆。
质粒分享者信息:武汉淼灵生物科技有限公司成立于2014年6月,是专业从事生物技术服务及生化试剂研发、生产和销售的公司。我们致力于研发出质优价适的精品试剂,打破国内企业在低端恶性竞争、国外公司漫天要价的痛心局面,不断开拓国内外市场,成为中国生物公司里的海尔和华为,国外生物公司中的ATCC和addgene。
- 质粒图谱1: png/1-1FG0105Sa02.png
质粒序列: 点击查看序列LOCUS Exported 8538 bp ds-DNA circular SYN 10-七月-2017
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pCDH-ECMV-MCS-3×FLAG-EF1-alpha-ZsGreen1-T2A-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8538)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2017年7月10日 from MiaoLingPlasmid
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8538
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 234..414
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 458..583
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1076..1309
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1801..1916
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 1974..2277
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2278..2481
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 2576..2614
/note="MCS"
CDS 2633..2704
/codon_start=1
/product="three tandem FLAG(R) epitope tags"
/note="3xFLAG"
/translation="DYKDDDDKDYKDDDDKDYKDDDDK"
promoter 2758..2969
/note="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2982..3250
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 3278..3970
/codon_start=1
/product="Zoanthus green fluorescent protein"
/note="ZsGreen1"
/note="mammalian codon-optimized"
/translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
AIASGSALP"
CDS 3980..4033
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/note="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/protein_id="
"
/translation="EGRGSLLTCGDVEENPGP"
CDS 4034..4633
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4634..5221
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5295..5528
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5600..5721
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 5740..5875
/note="SV40 ori"
/note="SV40 origin of replication"
rep_origin complement(6315..6903)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7074..7934)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7935..8039)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
1381 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
1681 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
1741 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc
1801 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
1861 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
1921 gtttattaca gggacagcag agatccagtt tatcgataag cttgggagtt ccgcgttaca
1981 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
2041 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
2101 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
2161 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
2221 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
2281 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
2341 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
2401 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
2461 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
2521 cgctgttttg acctccatag aagacaccga ctctactaga ggatctattt ccggtgaatt
2581 cctcgagact agttctagaa gcggccgcgg atcctcggta ccaagcttaa gtgactacaa
2641 ggatgacgat gacaaggatt acaaagacga cgatgataag gactataagg atgatgacga
2701 caaataaaga tccatcgata ctagtaagga tctgcgatcg ctccggtgcc cgtcagtggg
2761 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg
2821 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
2881 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
2941 ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt
3001 cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc
3061 ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt
3121 cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca
3181 cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc
3241 gttacagatc caagctgtga ccggcgccta ctctagcatg gcccagtcca agcacggcct
3301 gaccaaggag atgaccatga agtaccgcat ggagggctgc gtggacggcc acaagttcgt
3361 gatcaccggc gagggcatcg gctacccctt caagggcaag caggccatca acctgtgcgt
3421 ggtggagggc ggccccttgc ccttcgccga ggacatcttg tccgccgcct tcatgtacgg
3481 caaccgcgtg ttcaccgagt acccccagga catcgtcgac tacttcaaga actcctgccc
3541 cgccggctac acctgggacc gctccttcct gttcgaggac ggcgccgtgt gcatctgcaa
3601 cgccgacatc accgtgagcg tggaggagaa ctgcatgtac cacgagtcca agttctacgg
3661 cgtgaacttc cccgccgacg gccccgtgat gaagaagatg accgacaact gggagccctc
3721 ctgcgagaag atcatccccg tgcccaagca gggcatcttg aagggcgacg tgagcatgta
3781 cctgctgctg aaggacggtg gccgcttgcg ctgccagttc gacaccgtgt acaaggccaa
3841 gtccgtgccc cgcaagatgc ccgactggca cttcatccag cacaagctga cccgcgagga
3901 ccgcagcgac gccaagaacc agaagtggca cctgaccgag cacgccatcg cctccggctc
3961 cgccttgccc gcggccgctg agggcagagg aagtcttcta acatgcggtg acgtggagga
4021 gaatcccggc cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
4081 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
4141 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
4201 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
4261 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
4321 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
4381 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
4441 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
4501 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
4561 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
4621 gcccggtgcc tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
4681 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
4741 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
4801 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
4861 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
4921 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
4981 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
5041 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
5101 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
5161 tcttccgcgt ctccgccttc gccctcagac gagtcggatc tccctttggc cgcctccccg
5221 cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
5281 agaaaagggg ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc
5341 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
5401 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
5461 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
5521 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
5581 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
5641 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
5701 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
5761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
5821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac
5881 ttttgcagag acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
5941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
6001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
6061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
6121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
6181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
6241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
6301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
6361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
6421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
6481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
6541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
6601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
6661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
6721 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
6781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
6841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
6901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
6961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
7021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
7081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
7141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
7201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
7261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
7321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
7381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
7441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
7501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
7561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
7621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
7681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
7741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
7801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
7861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
7921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
7981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
8041 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
8101 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
8161 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
8221 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
8281 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
8341 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
8401 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
8461 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
8521 acggccagtg ccaagctg
//
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