| 质粒名称: | pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31476 |
| 存储实验室: | Andrew Scharenberg |
|
载体骨架基本信息 |
|
| 载体名称: | pCVL |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5092 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | SFFV d14 GFP donor EF1s I-SceI |
| 物种来源: | -- |
| 插入大小: | 2173 bp |
| 基因突变: | GFP has 14 amino acids deleted from c terminus |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
HA
NLS
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt) was a gift from Andrew Scharenberg (Addgene plasmid # 31476 ; http://n2t.net/addgene:31476 ; RRID:Addgene_31476) |
| 参考文献部分: | Tracking genome engineering outcome at individual DNA breakpoints. Certo MT, Ryu BY, Annis JE, Garibov M, Jarjour J, Rawlings DJ, Scharenberg AM. Nat Methods. 2011 Jul 10;8(8):671-6. doi: 10.1038/nmeth.1648. 10.1038/nmeth.1648 PubMed 21743461 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt) 7229 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCVL SFFV d14GFP EF1s HA.NLS.Sce(opt)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7229)
AUTHORS Certo MT, Ryu BY, Annis JE, Garibov M, Jarjour J, Rawlings DJ,
Scharenberg AM
TITLE Tracking genome engineering outcome at individual DNA breakpoints.
JOURNAL Nat Methods. 2011 Jul 10;8(8):671-6. doi: 10.1038/nmeth.1648.
PUBMED 21743461
REFERENCE 2 (bases 1 to 7229)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7229
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 9..235
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 236..416
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 463..588
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1080..1313
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 1809..1926
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1955..2361
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 2376..2385
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2382..2900
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN173
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIE"
primer_bind complement(2427..2448)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2688..2707)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3035..3056
/label=EGFP-C
/note="EGFP, forward primer"
promoter 3086..3297
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 3306..3315
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3321..3347
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 3357..3377
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
misc_feature 4104..4692
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4157..4177)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4575..4586)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
protein_bind 4768..4801
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind complement(5006..5023)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5012..5236
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5309..5326)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5480..6068)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5560..5579)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6239..7099)
/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 6862..6881
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7100..7204)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacgtcaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
901 aaaagtaaga ccaccgcaca gcaagcggcc ctgatcttca gacctggagg aggagatatg
961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
1381 aatgaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
1921 aattttatcg attacgcgtc acgtgctagc tgcagtaacg ccattttgca aggcatggaa
1981 aaataccaaa ccaagaatag agaagttcag atcaagggcg ggtacatgaa aatagctaac
2041 gttgggccaa acaggatatc tgcggtgagc agtttcggcc ccggcccggg gccaagaaca
2101 gatggtcacc gcagtttcgg ccccggcccg aggccaagaa cagatggtcc ccagatatgg
2161 cccaaccctc agcagtttct taagacccat cagatgtttc caggctcccc caaggacctg
2221 aaatgaccct gcgccttatt tgaattaacc aatcagcctg cttctcgctt ctgttcgcgc
2281 gcttctgctt cccgagctct ataaaagagc tcacaacccc tcactcggcg cgccagtcct
2341 ccgacagact gagtcgcccg gatcgatcct cgagcgccac catggtgagc aagggcgagg
2401 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
2461 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
2521 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
2581 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
2641 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
2701 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
2761 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
2821 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
2881 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
2941 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
3001 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgtgag
3061 gatccaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
3121 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
3181 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
3241 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggat
3301 ctgccgccac catgggcgta tacccctacg acgtgcccga ctacgccccc gggcccccta
3361 agaaaaagag gaaggtgaag aacatcaaga agaaccaggt catgaacctg ggccccaaca
3421 gcaagctgct gaaggagtac aagagccagc tgatcgagct gaacatcgag cagttcgagg
3481 ccggcatcgg cctgatcctg ggcgacgcct acatcaggag cagggacgag ggcaagacct
3541 actgcatgca gttcgagtgg aagaacaagg cctacatgga ccacgtgtgc ctgctgtacg
3601 accagtgggt gctgagcccc ccccacaaga aggagagggt gaaccacctg ggcaacctgg
3661 tcatcacctg gggcgcccag accttcaagc accaggcctt caacaagctg gccaacctgt
3721 tcatcgtgaa caacaagaag accatcccca acaacctggt ggagaactac ctgaccccca
3781 tgagcctggc ctactggttc atggacgacg gcggcaagtg ggactacaac aagaacagca
3841 ccaacaagag catcgtgctg aacacccaga gcttcacctt cgaggaggtg gagtacctgg
3901 tgaagggcct gaggaacaag ttccagctga actgctacgt gaagatcaac aagaacaagc
3961 ccatcatcta catcgacagc atgagctacc tgatcttcta caacctgatc aagccctacc
4021 tgatccccca gatgatgtac aagctgccca acaccatcag cagcgagacc ttcctgaagt
4081 gacctgcagg taatgcattc gacaatcaac ctctggatta caaaatttgt gaaagattga
4141 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
4201 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
4261 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
4321 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
4381 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
4441 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
4501 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
4561 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
4621 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
4681 ccgcctcccc gcctggaatt cggtaccttt ttaaaagaaa aggggggact ggaagggcta
4741 attcactccc aacgaagaca agatatcata acttcgtata gcatacatta tacgaagtta
4801 taatttattt gtgaaatttg tgatgctatt gctttatttg taaccatatg tttatttgtg
4861 aaatttgtga tgctattgct ttatttgtaa ccattgcttt ttgcttgtac tgggtctctc
4921 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
4981 cctcaataaa gcttgcctcg accagcctcg actgtgcctt ctagttgcca gccatctgtt
5041 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
5101 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
5161 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
5221 gcggtgggct ctatggcctg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
5281 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
5341 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
5401 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
5461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
5521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
5581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
5641 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
5701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
5761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
5821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
5881 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
5941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
6001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
6061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
6121 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
6181 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
6241 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
6301 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
6361 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
6421 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
6481 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
6541 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
6601 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
6661 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
6721 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
6781 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
6841 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
6901 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
6961 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
7021 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
7081 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
7141 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
7201 cgcgcacatt tccccgaaaa gtgccacct
//
