| 质粒名称: | pCfB8644 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 126919 |
| 存储实验室: | Irina Borodina |
|
载体骨架基本信息 |
|
| 载体名称: | pCfB6682 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 10708bp |
| 载体类型: | 酵母表达 |
| 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: | XdCrtI and HMG1 |
| 物种来源: | Xanthophyllomyces dendrorhous and Yarrowia lipolytica |
| 克隆方法: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pCfB8644 was a gift from Irina Borodina (Addgene plasmid # 126919 ; http://n2t.net/addgene:126919 ; RRID:Addgene_126919) |
| 参考文献部分: | A teaching protocol demonstrating the use of EasyClone and CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae and Yarrowia lipolytica. Milne N, Tramontin LRR, Borodina I. FEMS Yeast Res. 2019 Sep 26. pii: foz062. doi: 10.1093/femsyr/foz062. 10.1093/femsyr/foz062 PubMed 31556952 |
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 pCfB8644 10708 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Plasmid containing integration cassette at IntC2 integration site in
Yarrowia lipolytica for overexpression of XdCrtI and XdCrtYB.
ACCESSION .
VERSION .
KEYWORDS pCfB8644
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10708)
AUTHORS Milne N, Tramontin LRR, Borodina I
TITLE A teaching protocol demonstrating the use of EasyClone and
CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae
and Yarrowia lipolytica.
JOURNAL FEMS Yeast Res. 2019 Sep 26. pii: foz062. doi:
10.1093/femsyr/foz062.
PUBMED 31556952
REFERENCE 2 (bases 1 to 10708)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10708
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 1..34
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
protein_bind complement(7965..7998)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
primer_bind complement(8468..8485)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8468..8484)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8583..8600)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8754..9342)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8834..8853)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9464..10324)
/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 10087..10106
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10325..10416)
/gene="bla"
/label=AmpR promoter
primer_bind 10458..10477
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 10483..10502
/label=pENTR-R
/note="pENTR vectors, reverse primer"
ORIGIN
1 gaagttccta ttctctagaa agtataggaa cttctgaagt ggggatttaa atgcggccgc
61 gaatgcacgc gatgattgaa acgcctgaca aaaacgcctg ataaaaacgc ctgataaaaa
121 cgcctgataa aaacgcctga taaaaacgcc tgtcaaaacc aaccagacct cggtagagag
181 gatcagagtc ccatggtgta ctatacagtg aaactgctgt gcgaatggta taccattgct
241 gtctatatag cctctagagt ggcactaatc cacaaggaca cagcacaatg tactacaggg
301 tagtactcac cccacagatg ctcttcgatg aaatagacag agaatgactg ggccagtacc
361 attgccaact tgtagggaat cgtcactctc caattccagc tccagttcca gttccagctc
421 tttcggctta attctagctc tcatttcggc atcttcattc agctgttcag gttcctgaca
481 ctgctcaatt cggcgatctc gacgactgca tcctgacttc agtctcgacc ctataattgt
541 gggctgcaca ctcggaggta taggccactc agatgcattc ttgggcggtc tataccagct
601 cattgaccct tcgtgggtct caatgttcaa cgcagatgaa cttcaaatgc ttatcagtgg
661 aggacatgcc aatgtggacg tctgggattt gaagacacac accaattacg gaggctatct
721 ggatacagat caaactataa aggacttctg gactgtgttt gagagtttcg aggaagagga
781 caagcggctg cttcttaagt ttgtgacatc agtatccaag gcaccattgc aaggattcaa
841 ggctttgaac ccgtcatttg ccattcgtaa cgctggtaga caggttgatc ggttccctac
901 ggcctccacc tgtgtcaatc ttctcaagct gcctgactat caggacattg atcaacttcg
961 gaagaaactt ttgtatgcca ttcgatcgca tgctggtttc gatttgtctt agaggaacgc
1021 atatacagta atcatagaga ataaacgata ttcatttatt aaagtagata gttgaggtag
1081 aagttgtaaa gagtgataaa tagcttagat accacagaca ccctcggtga cgaagtactg
1141 cagatggttt ccaatcacat tgacctgctg gagcagagtg ttaccggcag agcactgttt
1201 attgctctgg ccctggcaca tgacaacgtt ggagagagga gggtggatca ggggccagtc
1261 aataaagacc tcaccagagc agtgctggta accgtcccag aagggcactt gagggacgat
1321 atctcctcgg tgggtgattc ggtagagctt tcggtctttg gacaccttgg agacatcggg
1381 gttctcctgg ccaaagaaga gtttatcgac ccagttagca aagccagcgt taccgacaat
1441 gggctgacca agagtaacaa cgaggggatc gtggccgtta accttgaggt tgattccgaa
1501 cagaaggaat gcacgcgatt tactgccctt cccatccgct catgaccaca ctcaagactt
1561 tccgtactct cctccatccg gcaactgtcc ttctctctcc gacgtctcct ttccaaacga
1621 ctttgatctc tcggccgatc agtaggtagc tcgcgcaagc cgctcgcatt cccgcttgaa
1681 cctcggtagg aagtcggtca attcccttat aagaatgttt ggcaagatcc tctgcgtagg
1741 cgactaatgg aagcgagtac gtcttccatt cgaaccggaa gctttctgag gcgtttgaag
1801 atggtaatgt ggacgaagga gatagactga ggagtttgtc gaaatcttga ggccgaggtt
1861 ccgtccaatc agtcgggatc gcaagctttg attcatcccg aagaccaaag aatgagagtg
1921 gtaggtaaaa tctcccttct gttgcgtccc ctttaatgtc cctagcaatg ttcaccaact
1981 gaagggcagt tcccatctct cggcttgcca ctaacacagc ttctctttct tctatggtgg
2041 cagggacctg acttggtgca cttgcccaag agacatagac caatagctcg gcgactgagc
2101 ctgctacaca tagaccatag tccagcaagt cagctgtggt ctcgataggc gtcttccgag
2161 cctggactgc ctctgtcgat aagggaaaga taagatcagt ggtgtatcct ctaaggagtt
2221 cgtcgagtgg gtatcgaggg atcagccctt gcaacttagc gagcaacctg aaggcgaaat
2281 ggtattgaac gggaaccctt tcggtaagga attgaacgag ctcggcaggc gagagcgaag
2341 gaggaggcgg gaggggatac attcccgtgg gtcgggaagg gtgcgaagga ggaagtaaag
2401 gcgaagaaag gatcttgtca ggttgcgaag ggtgtagcgg gggcccaaat agtagggtaa
2461 gaaaatcgga gaccatgtca attgtggcat gcgggttgga agatacttca ggagagtcga
2521 taagatcatc agtcacccgg cagaatgcgt atagtccaac cagcctctcc ctaacttcgc
2581 taggaaatcc agccgaggca acaaaaaagc tccggctctt tttctccaac aacttgactg
2641 ccagttccaa gtcacgtttt ggctgagaag agtatggtcg gctgctaaaa aacagggaga
2701 gcacaggcgg tgtaatgagg ggaaatgaag atggcatctt tttgttgcca taaatagttc
2761 gaccgtgtag caggtatagg gcctgagtat gatcgcaggc agacagaccc agaacaatca
2821 ttagattcgt cagtaagaag aacatagctt cctcaatggg tagtacacct ccaagcctcc
2881 accctacaat cttctcatcg ttgatcgacc aagagtcttg accgacagca acataatcta
2941 cccaaatcag atacaccgtc gggatcatga ttgttgcaat agttgacttt gctcggccac
3001 ttttccaatc gaaagcatat tcgcctgata atgctgccaa gagcatggta ggtggggtga
3061 tgagtaagga gagtgcccgc atgtagaagt agtgatctgt cacgagcggg tcgggcgatg
3121 ggctggggtg agcggtaaat aggtagataa tgggcagagg gatgagcgcc ttgagcgcga
3181 gagaaagggc ggacgatcta gtcttgggaa gcgcgagaga tgggagaagg tgcctagttg
3241 ccaagacgta gaccaagccg gtgattacgg tttgaatgac aaagaaagcg tactcttcat
3301 atggaacatc tagaaacgtt ccaaacacgc cttggccact ctccgctgat ggatatgtcc
3361 atgcgccatt tctgatgatc catgagtccc atggtgtggt tgcactaaac gcaataaata
3421 cgaggatcga tattttgtag atgtcaaatt ttgtcaaaat cggggaagtg agcaggccga
3481 gaagaccaag aattgggaga gtatagatca gatggatctg gtaatatgcg agagccgtca
3541 tatgacagat tgtgtgatgt gtagtttaga tttcgaatct gtggggaaag aaaggaaaaa
3601 agagactggc aaccgattgg gagagccact gtttatatat accctagaca agccccccgc
3661 ttgtaagatg ttggtcaatg taaaccagta ttaaggttgg caagtgcagg agaagcaagg
3721 tgtgggtacc gagcaatgga aatgtgcgga aggcaaaaaa atgaggccac ggcctattgt
3781 cggggctata tccagggggc gattgaagta cactaacatg acatgtgtcc acagaccctc
3841 aatctggcct gatgagccaa atccatacgc gctttcgcag ctctaaaggc tataacaagt
3901 cacaccaccc tgctcgacct cagcgccttc actttttgct aagacaaact gtacacgctg
3961 ttccagcgtt ttctgcctgc acctggtggg acatttggtg caacctaaag tgctcggaac
4021 ctctgtggtg tccagatcag cgcagcagtt ccgaggtagt tttgaggccc ttagatgatg
4081 caatggtgtc agtcgctgga tcacgagtct taatggcagt attcgttctt atttgtgcca
4141 ttgagccccg ttatcctcgt atcttctacc ccccatccca tccctttgtt ggtgcaaccc
4201 tacccattta ttgttgggtg cagcccaacc gacgtggaga gcttggcttg gccatataaa
4261 aaggcccccc cctagtggca atggcagaaa gtcagctgtg agttgttgaa tttgtcatct
4321 aggcggcctg gccgtcttct ccggggcaat tgttcctcta tagtactgcg tacactgtta
4381 tcagtagctg acgcagtagg atgtcctgca cgggtctttt tgtggggtgt ggagaaaggg
4441 gtgcttggag atggaagccg gtagaaccgg gctgcttggg gggatttggg gccgctgggc
4501 tccaaagagg ggtaggcatt tcgttggggt tacgtaattg cggcatttgg gtcctgcgcg
4561 catgtcccat tggtcagaat tagtccggat aggagactta tcagccaatc acagcgccgg
4621 atccacctgt aggttgggtt gggtgggagc acccctccac agagtagagt caaacagcag
4681 cagcaacatg atagttgggg gtgtgcgtgt taaaggaaaa aaaaagaagc ttgggttata
4741 ttcccgctct atttagaggt tgcgggatag acgccgacgg agggcaatgg cgccatggaa
4801 ccttgcggat atcgatacgc cgcggcggac tgcgtccgaa ccagctccag cagcgttttt
4861 tccgggccat tgagccgact gcgaccccgc caacgtgtct tggcccacgc actcatgtca
4921 tgttggtgtt gggaggccac tttttaagta gcacaaggca cctagctcgc agcaaggtgt
4981 ccgaaccaaa gaagcggctg cagtggtgca aacggggcgg aaacggcggg aaaaagccac
5041 gggggcacga attgaggcac gccctcgaat ttgagacgag tcacggcccc attcgcccgc
5101 gcaatggctc gccaacgccc ggtcttttgc accacatcag gttaccccaa gccaaacctt
5161 tgtgttaaaa agcttaacat attataccga acgtaggttt gggcgggctt gctccgtctg
5221 tccaaggcaa catttatata agggtctgca tcgccggctc aattgaatct tttttcttct
5281 tctcttctct atattcattc ttgaattaaa cacacatcaa cagtgcaggt atgggaaaag
5341 aacaagatca ggataaaccc acagctatca tcgtgggatg tggtatcggt ggaatcgcca
5401 ctgccgctcg tcttgctaaa gaaggtttcc aggtcacggt gttcgagaag aacgactact
5461 ccggaggtcg atgctcttta atcgagcgag atggttatcg attcgatcag gggcccagtt
5521 tgctgctctt gccagatctc ttcaagcaga cattcgaaga tttgggggag aagatggaag
5581 attgggtcga tctcatcaag tgtgaaccca actatgtttg ccacttccac gatgaagaga
5641 ctttcacttt ttcaaccgac atggcgttgc tcaagcggga agtcgagcgt tttgaaggca
5701 aagatggatt tgatcggttc ttgtcgttta tccaagaagc ccacagacat tacgagcttg
5761 ctgtcgttca cgtcctgcag aagaacttcc ctggcttcgc agcattctta cggctacagt
5821 tcattggcca aatcctggct cttcacccct tcgagtctat ctggacaaga gtttgtcgat
5881 atttcaagac cgacagatta cgaagagtct tctcgtttgc agtgatgtac atgggtcaaa
5941 gcccatacag tgcgcccgga acatattcct tgctccaata caccgaattg accgagggca
6001 tctggtatcc gagaggaggc ttttggcagg ttcctaatac tcttcttcag atcgtcaagc
6061 gcaacaatcc ctcagccaag ttcaatttca acgctccagt ttcccaggtt cttctctctc
6121 ctgccaagga ccgagcgact ggtgttcgac ttgaatccgg cgaggaacat cacgccgatg
6181 ttgtgattgt caatgctgac ctcgtttacg cctccgagca cttgattcct gacgatgcca
6241 gaaacaagat tggccaactg ggtgaagtca agagaagttg gtgggctgac ttagttggtg
6301 gaaagaagct caagggaagt tgcagtagtt tgagcttcta ctggagcatg gaccgaatcg
6361 tggacggtct gggcggacac aatatcttct tggccgagga cttcaaggga tcattcgaca
6421 caatcttcga ggagttgggt ctcccagccg atccttcctt ttacgtgaac gttccctcgc
6481 gaatcgatcc ttctgccgct cccgaaggca aagatgctat cgtcattctt gtgccgtgtg
6541 gccatatcga cgcttcgaac cctcaagatt acaacaagct tgttgctcgg gcaaggaagt
6601 ttgtgatcca aacgctttcc gccaagcttg gacttcccga ctttgaaaaa atgattgtgg
6661 cagagaaggt tcacgatgct ccctcttggg agaaagaatt taacctcaag gacggaagca
6721 tcttgggact ggctcacaac tttatgcaag ttcttggttt caggccgagc accagacatc
6781 ccaagtatga caagttgttc tttgtcgggg cttcgactca tcccggaact ggggttccca
6841 tcgtcttggc tggagccaag ttaactgcca accaagttct cgaatccttt gaccgatccc
6901 cagctccaga tcccaatatg tcactctccg taccatatgg aaaacctctc aaatcaaatg
6961 gaacgggtat cgattctcag gtccagctga agttcatgga tttggagaga tgggtatacc
7021 ttttggtgtt gttgattggg gccgtgatcg ctcgatccgt tggtgttctt gctttctgaa
7081 tcgcacgcat tccgttgaag tgtggatggg gaagtgagtg cccggttctg tgtgcacaat
7141 tggcaatcca agatggatgg attcaacaca gggatatagc gagctacgtg gtggtgcgag
7201 gatatagcaa cggatattta tgtttgacac ttgagaatgt acgatacaag cattgtccaa
7261 gtacaatact aaacatactg tacatactca tactcgtacc cgggcaacgg tttcacttga
7321 gtgcagtggc tagtgccctt actcgtacag tgtgcaatac tgcgtatcat agtctttgat
7381 gtatatcgta ttcattcatg ttagttgcgt aagcgttgca cgtttccatc taagacctac
7441 atttgtctgt tagctttgtg tagtcttaga cggcttgtag cagtccggag cgagattgtg
7501 tttctcgttg accataagct gaacactgct gtttccagcc tcagaagcgc agtcctcgta
7561 tcgtttctgc tgctgcggtt gttgttgttg ttgttgttgt tgttgttgtt ggatgtcgtc
7621 cttcgcgtct ctggcccatc accaaggtgg aaggtggatc ttcgtcttca cgaagttggt
7681 acttgttggc aaacatctca ttgaccatat cccacatcac cctggggcct gtgttcgaag
7741 gatagagcgg gtgcgcaacc acccagagaa gcattcaaaa caatacctac aagttgatct
7801 ctttcggtgt ctccagatac ccatctccac aagtgagctt tctccacgaa tcgtccattg
7861 acacctgtcc agaaacacca accggtattt caatgagtcg ccgagtgttc aggtcgtcga
7921 attatcgcac gcattcgcgg ccgcaaattt aaataaaatg aagtgaagtt cctatacttt
7981 ctagagaata ggaacttcta tagtgagtcg aataagggcg acacaaaatt tattctaaat
8041 gcataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
8101 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
8161 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
8221 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
8281 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
8341 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
8401 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
8461 gcttaagact ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca
8521 tccgtcaggg gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct
8581 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
8641 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
8701 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
8761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
8821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
8881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
8941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
9001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
9061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
9121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac
9181 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
9241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
9301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
9361 tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg
9421 tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc tgcttaccaa tgcttaatca
9481 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
9541 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagcgct gcgatgatac
9601 cgcgagaacc acgctcaccg gctccggatt tatcagcaat aaaccagcca gccggaaggg
9661 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
9721 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccatcgcta
9781 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
9841 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
9901 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
9961 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
10021 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
10081 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
10141 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
10201 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
10261 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
10321 tcatattctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
10381 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggtcagtgtt acaaccaatt
10441 aaccaattct gaacattatc gcgagcccat ttatacctga atatggctca taacacccct
10501 tgtttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
10561 aaacgccgta gcgccgatgg tagtgtgggg actccccatg cgagagtagg gaactgccag
10621 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgcccgg gctaattatg
10681 gggtgtcgcc cttattcgac tctatagt
//
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