| 质粒名称: | pGR595 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 213785 |
| 存储实验室: | Chang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 10016bp |
| 载体类型: | Yeast Expression, Synthetic Biology |
| 筛选标记: | Nourseothricin |
| 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: | BadBoy2 TP-DNAP1 |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 3000 bp |
|
文献引用方法 |
|
| 材料和方法部分: | pGR595 was a gift from Chang Liu (Addgene plasmid # 213785 ; http://n2t.net/addgene:213785 ; RRID:Addgene_213785) |
买家导航
pGR595载体质粒基本信息
pGR595质粒图谱载体图谱和pGR595载体序列质粒序列多克隆位点信息
pGR595质粒载体简介
pGR595质粒序列载体序列
LOCUS pGR595 10016 bp DNA circular SYN 14-APR-2025
DEFINITION Integrates at the CAN1 locus the BadBoy2 TP-DNAP1 variant and the
Nourseothricin resistance marker. Includes an I-SceI transient
expression cassette..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10016)
TITLE Continuous evolution of user-defined genes at 1-million-times the
genomic mutation rate
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10016)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT Sequence Label: pGR595
FEATURES Location/Qualifiers
source 1..10016
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(41..60)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 258..924
/gene="S. cerevisiae TDH3"
/label=GAP promoter
/note="promoter for glyceraldehyde-3-phosphate
dehydrogenase; also known as the TDH3 promoter"
primer_bind 794..817
/label=GPDpro-F
/note="S. cerevisiae GPD promoter, forward primer"
terminator 6033..6198
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(6088..6112)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
gene 6199..7318
/label=natMX6
/note="yeast selectable marker conferring nourseothricin
resistance"
promoter 6199..6542
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 6543..7115
/codon_start=1
/gene="Streptomyces noursei nat1"
/product="nourseothricin acetyltransferase"
/label=NrsR
/note="confers resistance to nourseothricin"
/translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
LCGLDTALYDGTASDGEQALYMSMPCP"
terminator 7121..7318
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
primer_bind complement(7856..7872)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7856..7872)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7869..7891)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7880..7896
/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(join(7904..7910,7911..7928,7929..7934))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7949..7970
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8087..8104)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8258..8846)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8338..8357)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(9017..9808,9809..9877))
/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 9640..9659
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9878..9982)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
61 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
121 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
181 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tggtgagtga
241 attccagttc gagtttatca ttatcaatac tgccatttca aagaatacgt aaataattaa
301 tagtagtgat tttcctaact ttatttagtc aaaaaattag ccttttaatt ctgctgtaac
361 ccgtacatgc ccaaaatagg gggcgggtta cacagaatat ataacatcgt aggtgtctgg
421 gtgaacagtt tattcctggc atccactaaa tataatggag cccgcttttt aagctggcat
481 ccagaaaaaa aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat
541 aggtccattc tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca
601 caacctcaat ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc
661 acgcatgtat ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg
721 aaaaagctga aaaaaaaggt tgaaaccagt tccctgaaat tattccccta cttgactaat
781 aagtatataa agacggtagg tattgattgt aattctgtaa atctatttct taaacttctt
841 aaattctact tttatagtta gtcttttttt tagttttaaa acaccaagaa cttagtttcg
901 aataaacaca cataaacaaa caaaatgaag aacatcaaga agaaccaagt tatgaacttg
961 ggtccaaact ctaagttgtt gaaggaatac aagtctcaat tgatcgaatt gaacatcgaa
1021 caattcgaag ctggtatcgg tttgatcttg ggtgacgctt acatccgatc tcgcgacgaa
1081 ggtaagacct actgtatgca attcgaatgg aagaacaagg cttacatgga ccacgtttgt
1141 ttgttgtacg accaatgggt tttgtctcca ccacacaaga aggaaagagt taaccacttg
1201 ggtaacttgg ttatcacctg gggtgctcaa accttcaagc accaagcttt caacaagttg
1261 gctaacttgt tcatcgttaa caacaagaaa accatcccaa acaacttggt tgaaaactac
1321 ttgaccccaa tgtctttggc ttactggttc atggacgacg gtggtaagtg ggactacaac
1381 aagaactcta ccaacaagtc tatcgttttg aacacccaat ctttcacctt cgaagaagtt
1441 gaatacttgg ttaagggttt gagaaacaag ttccaattga actgttacgt taagatcaac
1501 aagaacaagc caatcatcta catcgactct atgtcttact tgatcttcta caacttgatc
1561 aagccatact tgatcccaca aatgatgtac aagttgccaa acaccatctc ttctgaaacc
1621 ttcttgaagt aaataaagca atcttgatga ggataatgat ttttttttga atatacataa
1681 atactaccgt ttttctgcta gattttgtga tgacgtaaat aagtacatat tactttttaa
1741 gccaagacaa gattaagcat taactttacc cttttctttc taagtttcaa tattagttat
1801 cactgtttaa aagttatggc gagaacgtcg gcggttaaaa tatattaccc tgaacggaat
1861 tctatatctt taacagattc caaaccctaa agtgtccgaa ttttcaatag ggcgaacttg
1921 aagaataacc aaggtcaata atatatcttt tagtataacc ctgaaatttg ccctatagaa
1981 atctagggtt tctgtgtggt ttccgggtga gtcatacggc ttttttgaat ttcttttttt
2041 gcagttgtct ctatcaatga aaatttcgag gaagacgata aggttaagat aagtagataa
2101 gagaatgata cgagataaag cacaaattag cagaaagaag agtggttgcg aacagagtaa
2161 accgaatcag ggaatccctt tttgcaaaaa catcaattat ccttttcttt tttttacgta
2221 tatatctgga acagaaatat ataagttact attatactta tagttggatc cagtttttaa
2281 tctgtcgtca atcgaaagtt tatttcagag ttcttcagac ttcttaactc ctgtaaaaac
2341 aaaaaaaaaa aaaggcatag catttgagaa tgaccttcca cgagctaaat tgaaagggaa
2401 gaatttatta gttgaactca agaaagaaga ggatgacgtg ggaaatggca tagaatccct
2461 tactaaatcg aacactaaac tgaactccat gctggcgaac gaaggtaaga tacacaaagc
2521 tagtttccag aaaagtgtaa aatttaaact acctgataat atagtgactg aagaaaccgt
2581 ggaacttaaa gaaataaagg acttgctact acaaatgttg agatgacagc gagagattga
2641 atcaagatta tccaatatcg aacttcaact cacggaaata ccgaaacata agtaatcata
2701 tcccttctca cattttttac acaggaagta agcaagttat gttatatttc cgacactata
2761 attaattctt agcagttaaa ggtgctttgt ctatattaca tttacataca gcttgagtga
2821 tcctgaccgg atatagggtc ctattttctt acgtgaacgg cttttcttct tgttcccgat
2881 ggccttcatg tgaaaaagca ctcctcggga ggcggaaaaa tatcaaaagt acggggcgaa
2941 gtttataatg aagatttatc gatataaatt ttggttattt caggagaaca agaaagctct
3001 ttacactaaa attatcagag aagaagctga tatattagcc ctaaggagta caccaaaaca
3061 caatggacta caaagacaaa gcattaaacg acttgagaaa cgtatacgca gacttcgact
3121 ccttgccatt ggacttcaga caaatcttaa ttaaggatag agctacattg ttgcaaaagg
3181 aagacgttga aaagaaaatt ttggaaagac aagaagatgc caaaaagtac gctgaatact
3241 tgaagcaatc agaaatccca gaaagaatct ccttacctaa catcaagaga cataagggtg
3301 tatcaatctc ctttgaagaa acatctgaag atatggtctt ggaaccaaga ccttttatct
3361 tcgacggttt gaacatcaga tgtttcagaa gagaaaccat attcagtttg aaaaataaga
3421 ttttgaacat ggttaaggaa tcttcatcct ttaaaaacgt ttcaagacaa agtgtatctt
3481 ttatgtactt caaaattttt aataagggta aagttattgc aagtacaaag tctgttaaca
3541 tctatgaaga taagatcgac gaaagattgg aagatttgtg taacaacttc gatgacgtat
3601 tgaaaaagat tatagacgtc acctatggtt acgaatcctt attcgttagt gaaacttact
3661 cttacgtaat attctatgct aaatcaatct actttccaca acctagatgc gttaacaact
3721 ggggtaacaa catcccaaac atcttgacat tcgattcctt taagcttttc accgcaaaca
3781 aaaacaacgt tagttgtatc aagcaatgtt caagattttt gtggcaaaaa gatttcaaca
3841 ctttagaaga aatgatagaa tacaagaacg gtaatatttg tatcgtcaca ccacaattgc
3901 atatcaacga tgttagagac atcaagtcat tcaacgatat cagattgtac tcagaatccc
3961 ctattaaaac attttccgtt attgataaca ctatcacata tttgttttac ttcaaggaac
4021 atttgggtgt tatttttaac atcactaagt caagacatga tagaagagta acaaaatttt
4081 caccattgtc caagttcagt gacgtcaaaa atatcactgt ttgtttcgat atcgaaagtt
4141 acttcgaccc agaaaaggaa tctaaccaag ttaacatccc tttcatttgt tgcgcttcaa
4201 tcatctataa taaggttatt ggtaacatcg tcgattttga aggtagagac tgtgtcgcac
4261 aaatgatcga atacgttgta gatatttgcg gtgaattgaa cataagttct gttgaattga
4321 tcgcccataa cggtggtggt tatgatttcc actacatctt gtcatccatg tataatccag
4381 ctgcaattaa aaacatattg atcagaaaca actcttttat ttcttttaat ttcgcacatg
4441 atggtgtcaa gttctctgtt aaggacagtt actcattttt gtcgtgtagt ttggccaacg
4501 cttctaaagc atttttaaac gaaggaactt ttaagaaaac tgatttccct catcacgact
4561 tgaaaactgc cgatgactta tataaggtct acaaggaatg gagttctgtt aacacagaaa
4621 ttaaccatgt cgttgaaaag gaaaagttgt tgatcacctc tgaacacatc gttaacttca
4681 ccaagaacga taagagtaag actttgatcg aatggtctaa ggattactgt agaaacgacg
4741 tattggtctt gtctaaggtt tggttggagt ttaaaaacgc tgtagaagat atttttaact
4801 gcgaattggt agaccaaacc atgactttgg ctggtttgtc atacaagttg ttccaagcaa
4861 acatgccatt cgatgtcgaa ttgagacatc ctaataagga agactacttc aacatgagag
4921 aagcattgat tggtggtaga tgtatctctg ttaacggtat ctacagggat gttttgtgct
4981 tagacgtaaa atcattatat ccagcttcta tggctttcta tgatcaacct tacggttctt
5041 ttaagagagt ttcatccaga ccaaaagatg aattgggtat ctattacgtt agagtaacca
5101 caaacagaaa caacaagtca aactttttcc ctatcagatc ccataataag atcacttaca
5161 acaacttcga agaatcaact tatattgctt ggtacacaaa cgttgatatt gacattggtt
5221 tgtcagaagg tcacaacatc gaatacatcc ctttcgattc ctatggtaac attggttact
5281 catggtccaa aaagggtaaa atttttgaaa agtacatcag ggatgttttg tacaaattga
5341 agataaaata tgaaaagcaa aataacaagg taaaaagaaa tgtcatcaaa aagatcatga
5401 actcattgtg gggtaaattc gcccaaaagt gggttaactt cgaatacttc atcaagtcag
5461 aagatgacat tgatttcgaa tccgaagaag catataaaat ttgggataca gacttcatgt
5521 tgatcaaaaa gattaaggaa tctatctaca gttctaagcc aatacaaaac ggtgttttta
5581 ctttaagttg ggccagatat cacatgaagt ctatttggga tgcaggtgcc aaagaaggtg
5641 ctgaatgtat ctattcagat acagactcca tattcgttca taaggaacac ttcaaaaaga
5701 atgcaaagtt tatgttgaac ggtttgaaag taccaatcat cggttctgaa gtcggtcaat
5761 ctgaattaga atgtgaattt gataaattgt tgtgcgccgg taaaaagcaa tacatgggtt
5821 tctacactta cttccaagat ggtaaacctt gcatcaagga aaagaaaaga ttcaaaggta
5881 taccaagtaa ctacatcata cctgaattgt atgctcactt gttatctggt gcagataagg
5941 aagccaaaat tcaatattta aaattcagaa gagaatgggg ttccgtcaag ggttacatag
6001 aaaataagac tgtcaaagcc acctaactcg aggcgaattt cttatgattt atgattttta
6061 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
6121 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
6181 tatagcatga ggtcgctcga catggaggcc cagaataccc tccttgacag tcttgacgtg
6241 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
6301 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
6361 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
6421 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
6481 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
6541 ccatgggtac cactcttgac gacacggctt accggtaccg caccagtgtc ccgggggacg
6601 ccgaggccat cgaggcactg gatgggtcct tcaccaccga caccgtcttc cgcgtcaccg
6661 ccaccgggga cggcttcacc ctgcgggagg tgccggtgga cccgcccctg accaaggtgt
6721 tccccgacga cgaatcggac gacgaatcgg acgacgggga ggacggcgac ccggactccc
6781 ggacgttcgt cgcgtacggg gacgacggcg acctggcggg cttcgtggtc gtctcgtact
6841 ccggctggaa ccgccggctg accgtcgagg acatcgaggt cgccccggag caccgggggc
6901 acggggtcgg gcgcgcgttg atggggctcg cgacggagtt cgcccgcgag cggggcgccg
6961 ggcacctctg gctggaggtc accaacgtca acgcaccggc gatccacgcg taccggcgga
7021 tggggttcac cctctgcggc ctggacaccg ccctgtacga cggcaccgcc tcggacggcg
7081 agcaggcgct ctacatgagc atgccctgcc cctaatcagt actgacaata aaaagattct
7141 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
7201 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
7261 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgat
7321 atgacgtttt attacctttg atcacatttc cacgccattt cgcattctca ccctcataag
7381 tcatacaccg aaaagaaagt ttaagggatc aatgagctta ctataatctc agtatattta
7441 tttttatcga tgattcacca caacaatctt gctcccgaaa agaaagcaga cggagtagaa
7501 gcatttgaaa ctccttcaga ccttcaagta tatatatata tatatatgta tatgtgtaca
7561 ttttcacgct aatactaatg tataattaga agataatttt tactcatttt tcgttatctt
7621 cacgtcaccc gaacctagaa ccaaatgtca ttttcacgat atgtaaatag tgaaataggc
7681 aaaaacgcca aaaagtagta agcgcaacat acactaaacc attaaagaat atctcgacca
7741 gaatctaaca gatatacatg ttccgataat gtctgagtta ggtgagtatt ctaaattaga
7801 aaacaaagag cttagaattc ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
7861 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
7921 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
7981 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
8041 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
8101 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
8161 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
8221 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
8281 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
8341 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
8401 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
8461 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
8521 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
8581 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
8641 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
8701 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
8761 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
8821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
8881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
8941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
9001 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
9061 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
9121 gcttaccatc tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag
9181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
9241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
9301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
9361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
9421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
9481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
9541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
9601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
9661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
9721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
9781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
9841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
9901 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
9961 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgac
//
