| 质粒名称: | pCDH-EF1-FHC |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 64874 |
| 存储实验室: | Richard Wood |
|
载体骨架基本信息 |
|
| 载体名称: | pCDH-CMV-MCS-EF1-Puro |
| 载体出品公司: | System Biosciences |
| 载体抗性: | 氨苄青霉素 |
| 载体修饰: | 添加FLAG和HA标记 |
| 载体类型: | 哺乳细胞表达,慢病毒 |
| 筛选标记: | 嘌呤霉素 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 标签/融合蛋白: |
Flag（主干上的C终端）
HA（主干上的C终端）
|
| 克隆方法: | 限制性内切酶 |
| 5' 测序引物 | EF1a-F_alt (gccgtgaacgttctttttc) |
| 3' 测序引物 | IRES-R (CCTCACATTGCCAAAAGACG) |
|
文献引用方法 |
|
| 材料和方法部分: | pCDH-EF1-FHC was a gift from Richard Wood (Addgene plasmid # 64874 ; http://n2t.net/addgene:64874 ; RRID:Addgene_64874) |
| 参考文献部分: | Mechanism of suppression of chromosomal instability by DNA polymerase POLQ. Yousefzadeh MJ, Wyatt DW, Takata K, Mu Y, Hensley SC, Tomida J, Bylund GO, Doublie S, Johansson E, Ramsden DA, McBride KM, Wood RD. PLoS Genet. 2014 Oct 2;10(10):e1004654. doi: 10.1371/journal.pgen.1004654. eCollection 2014 Oct. PGENETICS-D-14-01461 [pii] PubMed 25275444 |
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 pCDH-EF1-FHC 7714 bp ds-DNA circular SYN 23-MAR-2020
DEFINITION lentiviral vector with Flag, HA tags.
ACCESSION .
VERSION .
KEYWORDS pCDH-EF1-FHC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7714)
AUTHORS Yousefzadeh MJ, Wyatt DW, Takata K, Mu Y, Hensley SC, Tomida J,
Bylund GO, Doublie S, Johansson E, Ramsden DA, McBride KM, Wood RD
TITLE Mechanism of suppression of chromosomal instability by DNA
polymerase POLQ.
JOURNAL PLoS Genet. 2014 Oct 2;10(10):e1004654. doi:
10.1371/journal.pgen.1004654. eCollection 2014 Oct.
PUBMED 25275444
REFERENCE 2 (bases 1 to 7714)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 23, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7714
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 455..571
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 614..825
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 838..1106
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 1178..1201
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1217..1243
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_feature 1254..1826
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1421..1438)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 1648..1667
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 1835..2434
/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(1835..1854)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2331..2351
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 2441..3029
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2494..2514)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2912..2923)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3103..3336
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 3408..3529
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3445..3464)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3499..3518
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3569..3704
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3631..3650
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3737..3753)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3737..3753)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3750..3772)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3761..3777
/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(3785..3815)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3830..3851
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3968..3985)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4139..4727)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4219..4238)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4898..5758)
/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 5521..5540
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5759..5863)
/gene="bla"
/label=AmpR promoter
primer_bind 5931..5949
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5987..6009)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6109..6128
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6322..6344
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6336..6353
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6337..6353
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 6368..6594
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6595..6775
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6822..6947
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7440..7673
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
ORIGIN
1 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
61 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
121 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
181 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
241 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
301 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
361 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
421 gattagtgaa cggatctcga cggtatcggt taacttttaa aagaaaaggg gggattgggg
481 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
541 tacaaaaaca aattacaaaa ttcaaaattt tatcgatact agtggatctg cgatcgctcc
601 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
661 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
721 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
781 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
841 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
901 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
961 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
1021 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
1081 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctactct agagctagcg
1141 aattcgaatt taaatcggat ccgcggccgc tggaggagac tacaaggacg acgatgacaa
1201 gtcggccgct ggaggatacc cctacgacgt gcccgactac gcctagggcc gcgcccctct
1261 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
1321 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
1381 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
1441 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
1501 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
1561 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
1621 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
1681 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
1741 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
1801 gttttccttt gaaaaacacg ataatacctc cggaatgacc gagtacaagc ccacggtgcg
1861 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
1921 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
1981 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
2041 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
2101 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
2161 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
2221 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
2281 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
2341 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
2401 cacctggtgc atgacccgca agcccggtgc ctaggtcgac aatcaacctc tggattacaa
2461 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
2521 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
2581 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
2641 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
2701 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
2761 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
2821 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
2881 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
2941 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
3001 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
3061 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
3121 cccaacgaag ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
3181 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
3241 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
3301 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
3361 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
3421 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
3481 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
3541 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
3601 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
3661 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
3721 ccaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
3781 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
3841 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
3901 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
3961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
4021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
4081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
4141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
4201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
4261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
4321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
4381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
4441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
4501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
4561 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
4621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
4681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
4741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
4801 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
4861 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
4921 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
4981 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
5041 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
5101 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
5161 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
5221 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
5281 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
5341 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
5401 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
5461 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
5521 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
5581 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
5641 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
5701 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
5761 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
5821 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
5881 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
5941 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
6001 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
6061 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
6121 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
6181 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
6241 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
6301 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
6361 tgacgcgtgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
6421 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
6481 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
6541 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
6601 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
6661 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
6721 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
6781 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
6841 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
6901 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
6961 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
7021 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
7081 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
7141 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
7201 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
7261 aaagtaagac caccgcacag caagcggcca ctgatcttca gacctggagg aggagatatg
7321 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
7381 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
7441 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
7501 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
7561 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
7621 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
7681 tggggttgct ctggaaaact catttgcacc actg
//
