| 质粒名称: | CTCF-mAC donor (Hygro) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140646 |
| 存储实验室: | Masato Kanemaki |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达, CRISPR |
| 筛选标记: | 潮霉素 |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | CTCF |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | CTCF-mAC donor (Hygro) was a gift from Masato Kanemaki (Addgene plasmid # 140646 ; http://n2t.net/addgene:140646 ; RRID:Addgene_140646) |
| 参考文献部分: | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, Hayashi KI, Kanemaki MT. Nat Commun. 2020 Nov 11;11(1):5701. doi: 10.1038/s41467-020-19532-z. 10.1038/s41467-020-19532-z PubMed 33177522 |
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 CTCF-mAC donor (Hygro) 7263 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CTCF tagging with mAID-clover.
ACCESSION .
VERSION .
KEYWORDS CTCF-mAC donor (Hygro)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7263)
AUTHORS Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano
R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y,
Hayashi KI, Kanemaki MT.
TITLE The auxin-inducible degron 2 technology provides sharp degradation
control in yeast, mammalian cells, and mice.
JOURNAL Nat Commun (2020) 11(1):5701
REFERENCE 2 (bases 1 to 7263)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7263
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 585..607
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 599..616
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1115..1318
/codon_start=1
/product="minimal degron from the auxin-responsive IAA17
protein of Arabidopsis thaliana (Kubota et al., 2013)"
/label=mini-AID
/note="acts as an auxin-inducible degron (AID) in the
presence of TIR1"
/translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
AAFVKVSMDGAPYLRKIDLRMYK"
CDS 1325..2044
/codon_start=1
/product="bright green-yellow fluorescent protein derived
from mClover2, with improved photostability (Bajar et al.,
2003)"
/label=mClover3
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(1370..1391)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 1978..1999
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2162..2283
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2199..2218)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2253..2272
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
protein_bind complement(2314..2347)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 2408..2907
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(2443..2466)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 2840..2859
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 2968..3987
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
protein_bind complement(4550..4583)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind complement(5075..5095)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5075..5093)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5114..5130)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5114..5130)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5127..5149)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5138..5154
/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(5162..5192)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5207..5228
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5345..5362)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5516..6104)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5596..5615)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6275..7135)
/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 6898..6917
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7136..7240)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctctg
661 aggcaggaga attgctttaa cccggggggc agaggttgca gtgagctgag atcatgcccg
721 tgcactccag cctggtcata gagcaagact ccgtctcaaa aaataaaata aaaaataaat
781 tgactgtctc tggaccgcta tctaataagg ctgtcctgca ttgctgacat cccgttcgct
841 gtcagtctaa aagacccttg tgattcttgg ggctttaatg gaccatttgt tctgtctgtg
901 ctcttctttg ccagcaacag ctatcattca ggttgaagac cagaatacag gtgcaattga
961 gaacattata gttgaagtaa aaaaagagcc agatgctgag cccgcagagg gagaggaaga
1021 ggaggcccag ccagctgcca cagatgcccc caacggagac ctcacgcccg agatgatcct
1081 cagcatgatg gaccggggat ccggtgcagg cgccaaggag aagagtgctt gtcctaaaga
1141 tccagccaaa cctccggcca aggcacaagt tgtgggatgg ccaccggtga gatcataccg
1201 gaagaacgtg atggtttcct gccaaaaatc aagcggtggc ccggaggcgg cggcgttcgt
1261 gaaggtatca atggacggag caccgtactt gaggaaaatc gatttgagga tgtataaagc
1321 tagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
1381 ggacggcgac gtaaacggcc acaagttcag cgtccgcggc gagggcgagg gcgatgccac
1441 caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
1501 caccctcgtg accaccttcg gctacggcgt ggcctgcttc agccgctacc ccgaccacat
1561 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
1621 ctctttcaag gacgacggta cctacaagac ccgcgccgag gtgaagttcg agggcgacac
1681 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
1741 gcacaagctg gagtacaact tcaacagcca caacgtctat atcacggccg acaagcagaa
1801 gaacggcatc aaggctaact tcaagatccg ccacaacgtt gaggacggca gcgtgcagct
1861 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
1921 ccactacctg agccatcagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
1981 ggtcctgctg gagttcgtga ccgccgccgg gattacacat ggcatggacg agctgtacaa
2041 gtaatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
2101 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
2161 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
2221 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
2281 ttaacgcgtc gatcatattc aataaccctt aatataactt cgtataatgt atgctatacg
2341 aagttattag gtctgaagag gagtttacgt ccagccaagc ttaggatctc gacctcgaaa
2401 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
2461 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
2521 ggtaggcgcc aaccgactcc gttctttggt ggccccttcg cgccaccttc tactcctccc
2581 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
2641 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
2701 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
2761 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
2821 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
2881 tcctcttcct catctccggg cctttcgacc tgcatccatc tagatctcga gcagctgaag
2941 cttaccgcta gcatggatag atccggaaag cctgaactca ccgcgacgtc tgtcgagaag
3001 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
3061 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
3121 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
3181 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
3241 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
3301 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
3361 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
3421 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
3481 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
3541 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
3601 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
3661 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
3721 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
3781 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
3841 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
3901 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
3961 agcactcgtc cgagggcaaa ggaataggct agccgcccgc cccacgaccc gcagcgcccg
4021 accgaaagga gcgcacgacc ccatgcatcg atgatatcag atccccggga tgcagaaatt
4081 gatgatctat taaacaataa agatgtccac taaaatggaa gtttttcctg tcatactttg
4141 ttaagaaggg tgagaacaga gtacctacat tttgaatgga aggattggag ctacgggggt
4201 gggggtgggg tgggattaga taaatgcctg ctctttactg aaggctcttt actattgctt
4261 tatgataatg tttcatagtt ggatatcata atttaaacaa gcaaaaccaa attaagggcc
4321 agctcattcc tcccactcat gatctataga tctatagatc tctcgtggga tcattgtttt
4381 tctcttgatt cccactttgt ggttctaagt actgtggttt ccaaatgtgt cagtttcata
4441 gcctgaagaa cgagatcagc agcctctgtt ccacatacac ttcattctca gtattgtttt
4501 gccaagttct aattccatca gaagctggtc gagatccgga acccttaata taacttcgta
4561 taatgtatgc tatacgaagt tattaggtcc ctcgaagagg ttcactagga tccgcgtcgc
4621 caggacttct ctgggctgtg tttaaacggc ccgcatctta atttttctcc cttctttctt
4681 tttttggctt tgggaaaagc atcattttac caaacatacc gagaacgaaa acttcaagga
4741 tgatgttaga aaaaaatgtg atttaactag aacttgctgt ctgatgttag caaatcatgg
4801 aatgttctga gtccctgagg gtttactgtg aagtgctgag gacagtgttg acaactaact
4861 cgttttccta gatggaaacg gagacattga cccctccctc catgtggtaa accactccag
4921 aatggccacc aggcttccca gagttctatg gtcttcttcc caagagagtt tttaattgta
4981 aatgcatact tgggaaggac ttagagtttt aaactgtttt ttgcttttgc ttttccctga
5041 ctccctttgc ttggagggta cccagctttt gttcccttta gtgagggtta attgcgcgct
5101 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
5161 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
5221 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
5281 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
5341 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
5401 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
5461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
5521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
5581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
5641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
5701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
5761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
5821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
5881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
5941 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
6001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
6061 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
6121 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
6181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
6241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
6301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
6361 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
6421 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
6481 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
6541 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
6601 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
6661 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
6721 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
6781 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
6841 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
6901 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
6961 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
7021 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
7081 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
7141 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
7201 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
7261 cac
//
