| 质粒名称: | GFP-hChibby1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89742 |
| 存储实验室: | Ken-Ichi Takemaru |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+ |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 5800 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | human Chibby1 |
| 物种来源: | H. sapiens (human) |
| 标签/融合蛋白: | GFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | XhoI (未破坏) |
| 5’测序引物 | ACCACATGGTCCTTCTTCAG |
| 3’测序引物 | TTATGCTGAGTGATATC |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-hChibby1 was a gift from Ken-Ichi Takemaru (Addgene plasmid # 89472 ; http://n2t.net/addgene:89472 ; RRID:Addgene_89472) |
| 参考文献部分: | |
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 GFP-hChibby1 5843 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Labels mother centrioles/basal bodies.
ACCESSION .
VERSION .
KEYWORDS GFP-hChibby1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5843)
TITLE GFP-hChibby1
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5843)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5843
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 27..1004
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1039..1057
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1039..1056
/label=SP6
/note="SP6 promoter, forward primer"
CDS 1092..1805
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(1122..1150)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 1745..1765
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
primer_bind complement(2875..2894)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2876..2894)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2899..3033
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2923..2942)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2977..2996
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(3146..3166)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(3146..3164)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3185..3201)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3185..3201)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3198..3220)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3209..3225
/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(3233..3263)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3278..3299
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3416..3433)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3587..4175)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3667..3686)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4346..5206)
/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 4969..4988
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5207..5311)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5337..5792)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5474..5495)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5686..5705
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
61 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
121 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
181 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
241 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
301 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
361 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
421 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
481 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
541 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
601 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
661 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
721 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
781 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
841 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
901 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
961 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg
1021 gacgtcggag caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg
1081 caggatctac catgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
1141 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
1201 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
1261 ggccaacact tgtcactact ctcacttatg gtgttcaatg cttttcaaga tacccagatc
1321 atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
1381 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
1441 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
1501 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
1561 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
1621 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
1681 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc
1741 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
1801 acaaaggatc ccatcgattc gaattcatgc ctttctttgg gaatacgttc agtccgaaga
1861 agacacctcc tcggaagtcg gcatctctct ccaacctgca ttctttggat cgatcaaccc
1921 gggaggtgga gctgggcttg gaatacggat ccccgactat gaacctggca gggcaaagcc
1981 tgaagtttga aaatggccag tggatagcag agacaggggt tagtggcggt gtggaccgga
2041 gggaggttca gcgccttcgc aggcggaacc agcagttgga ggaagagaac aatctcttgc
2101 ggctgaaagt ggacatctta ttagacatgc tttcagagtc cactgccgaa tcccacttaa
2161 tggagaagga actggatgaa ctgaggatca gccggaagag aaaatgaaga ccccagagac
2221 atttattggg gagtaggatg tggctgagtg cttttttttt tggccagact agcggattca
2281 gtcctggaag agagtatcat ataatgagac ccacaggcac tggcaccctt gggttggcaa
2341 tagaaggtga catggaatgg agaaaaccaa gattccagat ggggatagta actagaaggt
2401 gcttcagatg cactgcctgc gggtgccagt ctgaaaacca gaccgcacag aggcctgggg
2461 ctgctgatga gcttttcggt gctctccaca cacaagctcg caaacacaca tgtcccagaa
2521 tagctctgtt gggttgtgtt gggagaagcg gctggagttc attctctcac ccccttatgt
2581 tggtgtttgg cgtgtgacag cagttctaca gagctctgtg ttggggtcat ggatgagcgg
2641 ctctcttggc tcttaaaggc aggcctctct cttcttgcct ttaaagaatc ctccttcctc
2701 acacctggcc tcctctggct tcagcttctc agcagcaagc accagccttc cacaacaaca
2761 ctatattttt atgctacttt cctgtttgca ctactacttt tttattaaac gatgttaaat
2821 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa rwmwmgmgcc tctagattct gcagccctat
2881 agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc
2941 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
3001 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
3061 tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat
3121 tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
3181 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
3241 gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
3301 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
3361 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
3421 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
3481 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
3541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
3601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
3661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
3721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
3781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
3841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
3901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
3961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
4021 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
4081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
4141 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
4201 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
4261 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
4321 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
4381 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
4441 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
4501 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
4561 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
4621 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
4681 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
4741 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
4801 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
4861 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
4921 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
4981 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
5041 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
5101 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
5161 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
5221 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
5281 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
5341 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
5401 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
5461 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
5521 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
5581 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
5641 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
5701 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
5761 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
5821 ttgggaaggg cgatcggtgc ggg
//
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