| 质粒名称: | pCVL Traffic Light Reporter 1.1 (Sce target) Ef1a BFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31481 |
| 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: | 5000 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | BFP fluorescense |
| 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: | Traffic Light Reporter 1.1 (Sce target) EF BFP |
| 物种来源: | -- |
| 插入大小: | 2908 bp |
| 基因突变: | mCherry含有M9S和M16L以减少来自内部起始密码子的背景荧光。这就是“.1”的含义。 |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 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 Traffic Light Reporter 1.1 (Sce target) Ef1a BFP was a gift from Andrew Scharenberg (Addgene plasmid # 31481 ; http://n2t.net/addgene:31481 ; RRID:Addgene_31481) |
| 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 |
pCVL Traffic Light Reporter 1.1 (Sce target) Ef1a BFP
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
 Ef1a BFP质粒图谱.png)
载体简介
载体序列
LOCUS pCVL Traffic Light Reporter 1.1 (Sce target) Ef1a BFP 7995 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCVL Traffic Light Reporter 1.1 (Sce target) Ef1a BFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7995)
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 7995)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7995
/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)"
primer_bind complement(2427..2448)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2709..2728)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3056..3077
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3134..3187
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 3188..3895
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNSAIIKEFLRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(3330..3348)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3598..3617
/label=mCherry-F
/note="mCherry, forward primer"
promoter 3922..4133
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 4152..4853
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
DLPSKLGHKLN"
misc_feature 4870..5458
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4923..4943)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5341..5352)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
protein_bind 5534..5567
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind complement(5772..5789)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5778..6002
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6075..6092)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6246..6834)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6326..6345)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7005..7865)
/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 7628..7647
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7866..7970)
/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 ggctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
2461 agttcagcgt gtccggcctc gacattaccc tgttatccct acgcaaataa gagctcacct
2521 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
2581 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
2641 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
2701 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
2761 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
2821 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
2881 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
2941 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
3001 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
3061 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
3121 aactggatcc ggtgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
3181 gggccctgtg agcaagggcg aggaggataa ctccgccatc atcaaggagt tcctgcgctt
3241 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
3301 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
3361 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
3421 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
3481 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctctctgca
3541 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
3601 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
3661 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
3721 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
3781 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
3841 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtgaatgca
3901 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
3961 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
4021 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
4081 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggatccac
4141 cggtcgccac catgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
4201 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
4261 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
4321 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
4381 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
4441 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
4501 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
4561 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
4621 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac atcaagacca
4681 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcgtctac tatgtggact
4741 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg
4801 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat tagtgaatct
4861 agagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
4921 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
4981 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
5041 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
5101 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
5161 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
5221 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
5281 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
5341 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
5401 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
5461 ggaattcggt acctttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
5521 aagacaagat atcataactt cgtatagcat acattatacg aagttataat ttatttgtga
5581 aatttgtgat gctattgctt tatttgtaac catatgttta tttgtgaaat ttgtgatgct
5641 attgctttat ttgtaaccat tgctttttgc ttgtactggg tctctctggt tagaccagat
5701 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
5761 gcctcgacca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
5821 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
5881 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
5941 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
6001 ggcctgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
6061 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
6121 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
6181 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
6241 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
6301 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
6361 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
6421 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
6481 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
6541 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
6601 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
6661 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
6721 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
6781 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
6841 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
6901 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
6961 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
7021 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
7081 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
7141 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
7201 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
7261 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
7321 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
7381 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
7441 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
7501 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
7561 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
7621 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
7681 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
7741 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
7801 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
7861 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
7921 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
7981 cgaaaagtgc cacct
//
