| 质粒名称: | pCY 3010-07 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 36212 |
| 存储实验室: | Anne Robinson |
|
载体骨架基本信息 |
|
| 载体名称: | pCY3010-02 |
| 载体抗性: | 氨苄西林 |
| 生长说明： | 用于DNA恢复的dh5alpha（即DNA预处理）。 |
| 载体类型: | 用于PCR和同源重组的酵母盒 |
| 筛选标记: | 博莱霉素 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | 红链霉菌博来霉素 |
| 物种来源: | 印度斯坦链异壁菌 |
| 插入大小: | 970 bp |
| 标签/融合蛋白: | yepolylinker cerulean zeocin（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | bglii（如果销毁则未知） |
| 3' 克隆位点: | SACI（如果销毁则未知） |
| 5' 测序引物 | CGAAAAGAGAGACCACATGGTCC |
|
文献引用方法 |
|
| 材料和方法部分: | pCY 3010-07 was a gift from Anne Robinson (Addgene plasmid # 36212 ; http://n2t.net/addgene:36212 ; RRID:Addgene_36212) |
| 参考文献部分: |
Cassette series designed for live-cell imaging of proteins and high-resolution techniques in yeast. Young CL, Raden DL, Caplan JL, Czymmek KJ, Robinson AS. Yeast. 2012 Mar;29(3-4):119-36. doi: 10.1002/yea.2895. Epub 2012 Apr 4. 10.1002/yea.2895 PubMed 22473760 |
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 pCY 3010-07 4237 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCY 3010-07
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4237)
AUTHORS Young CL, Raden DL, Caplan JL, Czymmek KJ, Robinson AS
TITLE Cassette series designed for live-cell imaging of proteins and
high-resolution techniques in yeast.
JOURNAL Yeast. 2012 Mar;29(3-4):119-36. doi: 10.1002/yea.2895. Epub 2012 Apr
4.
PUBMED 22473760
REFERENCE 2 (bases 1 to 4237)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4237
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(114..142)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 737..757
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
gene 854..1778
/label=bleMX6
/note="yeast selectable marker conferring phleomycin
resistance"
promoter 854..1197
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 1198..1572
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 1581..1778
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(1874..1893)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1875..1893)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1980..1997)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2151..2739)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2231..2250)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2910..3770)
/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 3533..3552
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3771..3875)
/gene="bla"
/label=AmpR promoter
primer_bind 3943..3961
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(3999..4021)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 4121..4140
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 4221..2
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 4221..1
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggcttaaga
61 tcggtgacgg tgctggttta attaacagta aaggagaaga acttttcact ggagttgtcc
121 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
181 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
241 tacctgttcc atggccaaca cttgtcacta ctttgacttg gggtgttcaa tgctttgcta
301 gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg
361 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
421 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
481 atggaaacat tcttggacac aaattggaat acaacgctat ttcagataat gtatacatca
541 ctgcagacaa acaaaagaat ggaatcaaag ctaatttcaa aattagacac aacattgaag
601 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
661 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
721 aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acactaggca
781 tggatgaact atacaaatag ggcgcgccag atctgtttag cttgcctcgt ccccgccggg
841 tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
901 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
961 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
1021 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
1081 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
1141 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
1201 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
1261 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
1321 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
1381 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
1441 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
1501 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
1561 gagcaggact gataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
1621 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
1681 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
1741 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
1801 gccatcccga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
1861 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
1921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
1981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
2041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
2101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
2161 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
2221 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
2281 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
2341 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
2401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
2461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
2521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
2581 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
2641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
2701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
2761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
2821 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
2881 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
2941 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
3001 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
3061 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
3121 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
3181 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
3241 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
3301 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
3361 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
3421 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
3481 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
3541 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
3601 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
3661 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
3721 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
3781 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
3841 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
3901 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
3961 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
4021 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
4081 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
4141 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
4201 ttatgtatca tacacatacg atttaggtga cactata
//
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