| 质粒名称: | pHR_A08_hrGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 84615 |
| 存储实验室: | Ian Wheeldon |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2623 bp |
| 完整质粒大小: | 8873 bp |
| 载体修饰: | -- |
| 载体类型: | 酵母表达，合成生物学 |
| 筛选标记: | URA3 |
| 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: | hrGFP |
| 物种来源: | -- |
| 插入大小: | 720 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | UAS1B8-TEF(136) |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BssHII(未破坏) |
| 3' 克隆位点: | NheI (未破坏) |
| 5' 测序引物 | TEF |
| 3' 测序引物 | CYC |
|
文献引用方法 |
|
| 材料和方法部分: | pHR_A08_hrGFP was a gift from Ian Wheeldon (Addgene plasmid # 84615 ; http://n2t.net/addgene:84615 ; RRID:Addgene_84615) |
| 参考文献部分: | Standardized markerless gene integration for pathway engineering in Yarrowia lipolytica. Schwartz C, Shabbir-Hussain M, Frogue K, Blenner M, Wheeldon I. ACS Synth Biol. 2016 Dec 19. 10.1021/acssynbio.6b00285 PubMed 27989123 |
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 pHR_A08_hrGFP 8873 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION HR donor to integrate UAS1B8TEF(136)-hrGFP-CYC1t into A08 locus.
ACCESSION .
VERSION .
KEYWORDS pHR_A08_hrGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8873)
AUTHORS Schwartz C, Shabbir-Hussain M, Frogue K, Blenner M, Wheeldon I
TITLE Standardized markerless gene integration for pathway engineering in
Yarrowia lipolytica.
JOURNAL ACS Synth Biol. 2016 Dec 19.
PUBMED 27989123
REFERENCE 2 (bases 1 to 8873)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8873
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 118..137
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 331..353
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 345..362
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 346..362
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 2720..3439
/codon_start=1
/product="humanized Renilla green fluorescent protein"
/label=hrGFP
/note="mammalian codon-optimized"
/translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
AQLTSLGKPLGSLHEWV"
terminator 3447..3694
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(3463..3481)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind complement(4719..4735)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4719..4735)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4732..4754)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4743..4759
/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(4767..4797)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4812..4833
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4950..4967)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5121..5709)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5201..5220)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5880..6740)
/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 6503..6522
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6741..6845)
/gene="bla"
/label=AmpR promoter
primer_bind 6913..6931
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(8869..18)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
61 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
121 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
181 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
361 gtgaattcga gctcggtacc cgcttttcgt agataatgga atacaaatgg atatccagag
421 tatacacatg gatagtatac actgacacga caattctgta tctctttatg ttaactactg
481 tgaggcgtta aatagagctt gatatataaa atgttacatt tcacagtctg aacttttgca
541 gattacctaa tttggtaaga tattaattat gaactgaaag ttgatggcat ccctaaattt
601 gatgaaaggg ggatcccccg ggttcgaagg taccaaggaa gcatgcggta ctgcataaaa
661 cccgcataca cgtgcataat acttgcataa cgatagcata taaactgcac gattgctgca
721 cttgacttta tatcaaactc aatatcagca ccacccgatt ctctgattat ttcatcaact
781 ataggagaga atgagttggt gataagacat cggtaccaat aaaaatcacg tgactattcc
841 acaggccttc tgttatatca tcagatgcct ttgaacattc tcttgtcgaa ctgagtccaa
901 tttcaacaac tctttccaca gtgcagtgcc cggttaggcc aattagggtt tggcaaaaat
961 cggcaatata agacagcaaa ctaaaattta tataaagccc atcgctgcct tgttggagcc
1021 tcgttttttc ccacaacaca agcttagcag taattgggtc ttctattctc aattgcacgc
1081 tccccccctc tccttacaat tcgctcactc tcccgactat ttaacaacgc acgacaaaca
1141 tagcttcaat tccccattat gattatctgg ttatcggtgg aggctaagat ggtgtagctt
1201 ctgcttgtcg agccgtctcg tacggcgcca agacgctgct ggtcgagttc aaggcgctgg
1261 gaggcacctg cgtcaacgtg ggctgtgccc aaaaagatca cgtggaacgc ctccgatctg
1321 gcgggccgaa tccgacaggc ccaggagtac ggcttctccg acgtggaccc caagtccgcc
1381 gacaactttg actggaccgg attcaaggcc aaacgagacg cctacgtcaa acgactcaat
1441 ggcatctacg aacgaaacct ccagaaggag ggcgtggagt acgtgtttgg ttgggccacc
1501 ctctacaagc aggagggtca ggagttcccc ctggtactgg ccaagagcga cgatggcaac
1561 gccaagttgt actccgcaaa gaagattaag attgccaccg gcggcaagcc cattctgccc
1621 gacgtgcctg gagccgagta cggcattgat tactagtggt acccgaattc ctgaggtgtc
1681 tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt tgtgtgtagt gtacgtacat
1741 tatcgagacc gttgttcccg cccacctcga tccggtctag actgaggtgt ctcacaagtg
1801 ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag tgtacgtaca ttatcgagac
1861 cgttgttccc gcccacctcg atccggggat ccctgaggtg tctcacaagt gccgtgcagt
1921 cccgccccca cttgcttctc tttgtgtgta gtgtacgtac attatcgaga ccgttgttcc
1981 cgcccacctc gatccgggtc gacctgaggt gtctcacaag tgccgtgcag tcccgccccc
2041 acttgcttct ctttgtgtgt agtgtacgta cattatcgag accgttgttc ccgcccacct
2101 cgatccggga gctcctgagg tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc
2161 tctttgtgtg tagtgtacgt acattatcga gaccgttgtt cccgcccacc tcgatccggt
2221 ctagactgag gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt
2281 gtagtgtacg tacattatcg agaccgttgt tcccgcccac ctcgatccgg ggatccctga
2341 ggtgtctcac aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac
2401 gtacattatc gagaccgttg ttcccgccca cctcgatccg ggtcgacctg aggtgtctca
2461 caagtgccgt gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat
2521 cgagaccgtt gttcccgccc acctcgatcc gggcatgcct gcagcctaga agcttttgtg
2581 gttgggactt tagccaaggg tataaaagac caccgtcccc gaattacctt tcctcttctt
2641 ttctctctct ccttgtcaac tcacacccga aatcgttaag catttccttc tgagtataag
2701 aatcattcaa aggcgcgcca tggtgagcaa gcagatcctg aagaacaccg gcctgcagga
2761 gatcatgagc ttcaaggtga acctggaggg cgtggtgaac aaccacgtgt tcaccatgga
2821 gggctgcggc aagggcaaca tcctgttcgg caaccagctg gtgcagatcc gcgtgaccaa
2881 gggcgccccc ctgcccttcg ccttcgacat cctgagcccc gccttccagt acggcaaccg
2941 caccttcacc aagtaccccg aggacatcag cgacttcttc atccagagct tccccgccgg
3001 cttcgtgtac gagcgcaccc tgcgctacga ggacggcggc ctggtggaga tccgcagcga
3061 catcaacctg atcgaggaga tgttcgtgta ccgcgtggag tacaagggcc gcaacttccc
3121 caacgacggc cccgtgatga agaagaccat caccggcctg cagcccagct tcgaggtggt
3181 gtacatgaac gacggcgtgc tggtgggcca ggtgatcctg gtgtaccgcc tgaacagcgg
3241 caagttctac agctgccaca tgcgcaccct gatgaagagc aagggcgtgg tgaaggactt
3301 ccccgagtac cacttcatcc agcaccgcct ggagaagacc tacgtggagg acggcggctt
3361 cgtggagcag cacgagaccg ccatcgccca gctgaccagc ctgggcaagc ccctgggcag
3421 cctgcacgag tgggtgtaag ctagcctcat gtaattagtt atgtcacgct tacattcacg
3481 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
3541 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
3601 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
3661 gaaggttttg ggacgctcga aggctttaat ttgccctagg cttctttgct ctcgagaccc
3721 agcctaagcg agtggcggtg gttggaggag gctacattga cgtggagctg gccggtgtct
3781 tccacggact caactccgaa accacccttt tctgtcgagg ccagacggtg ctccgagcgt
3841 tcgacatcat gatccaggac accatcaccg actactaaca agaagggcat caacgtgctc
3901 aagggctctg gcgtcaagaa gattgtcaag aaggataatg gcgagctcac ctacgggcag
3961 gatggcgccg agaatgatat cactctcgat tcgctcattt ggatattgga cgagagcctc
4021 tcaaggacac cctcaacctg agcgagtttg gcatcaagac caacaagcgg ggctacattg
4081 aggtcaacga gtgccagcaa tcgtctttcg acaacattta ttcgcttgga gacgtttgct
4141 gcaaggtcga gctgacccca tggctattgc tactggacga aatctgtcca accggctgtt
4201 tggtcccgct gagttcaagg atcagaagca ggactacacc gatgttcctt cggccgtatt
4261 ttcccacccc gaggttggct ccaacggcat cacggaggct gccgccaagg agcagtatgg
4321 tgaggacaac gtcaagatct acacctctaa gtttgtcgcc atgtactacg ccatgttgca
4381 ggagaaagcc cctaccgcct acaagctggt gtgtgtcgga aaggacgaga aggttgttgg
4441 tctgcacatt gttggcaccg actcttccga gattctgcag ggttttggcg tggccgttcg
4501 aatgggagct accaaggccg atttcgacaa tgttgtggct ctttgtccca cttctgccga
4561 ggagctggtg acaatgggat ataccgggag cgcggcgaga gatttctctg gtttgggcta
4621 tcgatattta atcaactaga ctccactgtt gctgtcacag ggtcagagaa gatactagag
4681 tacggaagtg cgttccgttg aagcttggcg taatcatggt catagctgtt tcctgtgtga
4741 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
4801 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
4861 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
4921 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
4981 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
5041 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
5101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
5161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
5221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
5281 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
5341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
5401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
5461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
5521 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
5581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
5641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
5701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
5761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
5821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
5881 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
5941 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
6001 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
6061 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
6121 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
6181 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
6241 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
6301 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
6361 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
6421 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
6481 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
6541 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
6601 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
6661 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
6721 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
6781 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
6841 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
6901 ttaacctata aaaataggcg tatcacgagg cccagatcct ctagagtcga agcggccgct
6961 atgtctgata aaaggatgta acataggcaa gctgctcgtg agtgttgagt acgaacctta
7021 gatccaaatc acccgcaccc acggatatac ttgcttgaat atacagtagt atgcggccgc
7081 ttcgacacca tatcatataa aactaacaat gcagtcgacg agtatctgtc tgactcgtca
7141 ttgccgcctt tggagtacga ctccaactat gagtgtgctt ggatcacttt gacgatacat
7201 tcttcgttgg aggctgtggg tctgacagct gcgttttcgg cgcggttggc cgacaacaat
7261 atcagctgca acgtcattgc tggctttcat catgatcaca tttttgtcgg caaaggcgac
7321 gcccagagag ccattgacgt tctttctaat ttggaccgat agccgtatag tccagtctat
7381 ctataagttc aactaactcg taactattac cataacatat acttcactgc cccagataag
7441 gttccgataa aaagttctgc agactaaatt tatttcagtc tcctcttcac caccaaaatg
7501 ccctcctacg aagctcgagc taacgtccac aagtccgcct ttgccgctcg agtgctcaag
7561 ctcgtggcag ccaagaaaac caacctgtgt gcttctctgg atgttaccac caccaaggag
7621 ctcattgagc ttgccgataa ggtcggacct tatgtgtgca tgatcaagac ccatatcgac
7681 atcattgacg acttcaccta cgccggcact gtgctccccc tcaaggaact tgctcttaag
7741 cacggtttct tcctgttcga ggacagaaag ttcgcagata ttggcaacac tgtcaagcac
7801 cagtacaaga acggtgtcta ccgaatcgcc gagtggtccg atatcaccaa cgcccacggt
7861 gtacccggaa ccggaatcat tgctggcctg cgagctggtg ccgaggaaac tgtctctgaa
7921 cagaagaagg aggacgtctc tgactacgag aactcccagt acaaggagtt cctggtcccc
7981 tctcccaacg agaagctggc cagaggtctg ctcatgctgg ccgagctgtc ttgcaagggc
8041 tctctggcca ctggcgagta ctccaagcag accattgagc ttgcccgatc cgaccccgag
8101 tttgtggttg gcttcattgc ccagaaccga cctaagggcg actctgagga ctggcttatt
8161 ctgacccccg gggtgggtct tgacgacaag ggagacgctc tcggacagca gtaccgaact
8221 gttgaggatg tcatgtctac cggaacggat atcataattg tcggccgagg tctgtacggc
8281 cagaaccgag atcctattga ggaggccaag cgataccaga aggctggctg ggaggcttac
8341 cagaagatta actgttagag gttagactat ggatatgtca tttaactgtg tatatagaga
8401 gcgtgcaagt atggagcgct tgttcagctt gtatgatggt cagacgacct gtctgatcga
8461 gtatgtatga tactgcacaa cctgtgtatc cgcatgatct gtccaatggg gcatgttgtt
8521 gtgtttctcg atacggagat gctgggtaca agtagctaat acgattgaac tacttatact
8581 tatatgaggc ttgaagaaag ctgacttgtg tatgacttat tctcaactac atccccagtc
8641 acaataccac cactgcacta ccactacacc aaaaccatga tcaaaccacc catggacttc
8701 ctggaggcag aagaacttgt tatggaaaag ctcaagagag agaagccaag atactatcaa
8761 gacatgtgtc gcaacttcaa ggaggaccaa gctctgtaca ccgagaaaca ggcctttgtc
8821 gacgatctgg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tct
//
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