| 质粒名称: | pGR481 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 213783 |
| 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 |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 6220bp |
| 载体类型: | Yeast Expression, Synthetic Biology |
| 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: | ura3* |
|
文献引用方法 |
|
| 材料和方法部分: | pGR481 was a gift from Chang Liu (Addgene plasmid # 213783 ; http://n2t.net/addgene:213783 ; RRID:Addgene_213783) |
买家导航
pGR481载体质粒基本信息
pGR481质粒图谱载体图谱和pGR481载体序列质粒序列多克隆位点信息
pGR481质粒载体简介
pGR481质粒序列载体序列
LOCUS pGR481 6220 bp DNA circular SYN 14-APR-2025
DEFINITION Integrates onto p1 a catalytically dead TRP5 and URA3 that can be
used for selection of transversion mutations.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6220)
TITLE Continuous evolution of user-defined genes at 1-million-times the
genomic mutation rate
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6220)
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"Unpublished"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: pGR481
FEATURES Location/Qualifiers
source 1..6220
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 119..138
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(357..461)
/gene="bla"
/label=AmpR promoter
primer_bind 529..547
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(585..607)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS 1480..2283
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MHKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRNFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
primer_bind complement(4786..4803)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4957..5545)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5037..5056)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5716..6220,1..287,288..356))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEVDKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
ORIGIN
1 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
61 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
121 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
181 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
241 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
301 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
361 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
421 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
481 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
541 acgaggccct ttcgtctcac gcgtttcggt gatgacggtg aaaacctctg acacatgcag
601 ctcccggaga cgagtactgg tgattataat accatttagg tgggttgggt tcttaactag
661 gatcatggcg gcagaatcaa tcaattgatg ttgaaccttc aatgtaggga attcgttctt
721 gatggtttcc tccacagttt ttctccataa tcttgaagag gccaaaacat tagctttatc
781 caaggaccaa ataggcaatg gtggctcatg ttgtagggcc atgaaagcgg ccattcttgt
841 gattctttgc acttctggaa cggtgtattg ttcactatcc caagcgacac catcaccatc
901 gtcttccttt ctcttaccaa agtaaatacc tcccactaat tctctgacaa caacgaagtc
961 agtaccttta gcaaattgtg gcttgattgg agataagtct aaaagagagt cggatgcaaa
1021 gttacatggt cttaagttgg cgtacaattg aagttcttta cggattttta gtaaaccttg
1081 ttcaggtcta acactaccgg taccccattt aggaccaccc acagcaccta acaaaacggc
1141 atcagccttc ttggaggctt ccagcgcctc atctggaagt ggaacacctg tagcatcgat
1201 agcagcacca ccaattaaat gattttcgaa atcgaacttg acattggaac gaacatcaga
1261 aatagcttta agaaccttaa tggcttcggc tgtgatttct tgaccaacgt ggtcacctgg
1321 caaaacgacg atcttcttag gaacgtgcat ttataatcat aaactattaa atcatattaa
1381 gaatcattca attgattttt gggatttgga ttatatagaa aagtatgacc taattgactc
1441 cggcgaaaaa tatatgaaag tttttataat aattataaaa tgcataaagc tacatataag
1501 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
1561 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
1621 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
1681 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
1741 ctcttcgaag acagaaactt tgctgacatt ggtaatacag tcaaattgca gtattctgcg
1801 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
1861 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
1921 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
1981 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
2041 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
2101 gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga
2161 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
2221 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
2281 taattaggca gatgggtctt cttcgaatct caaatcccaa cctatctttg gacctaattt
2341 cggcaagact tcagcgacac tttggacatc tttatcacct ctaccagaaa tattgataac
2401 caaatgttga tcaggcttca tcgtcttagc caattcgcaa gcgccataaa cagcatgaga
2461 agattccaaa gcgggaataa taccttctaa ttgagataat aatttaaagc caagcagagc
2521 ctgagcgtca gtagctgcaa tgaattgagc acggccagta gatttccaat atgccaattc
2581 tggaccgaca cctgggtagt ctaacccagc agaaacagaa tgagtatcat ggacttgacc
2641 atcactatct tgcaagacat aagtcttgac accatggaag acaccaggtc taccggcagt
2701 tagagtagca gagtggaact ttgtatctac accatcacca ccggcttcca cacccaataa
2761 cttaacggaa gtatcatgct cgaatggtga aaacatacct gtagagttgg aaccaccccc
2821 aacacatgca acaactgcgt caggtaattt accattgttc atggcagcaa actgttcctt
2881 ggtttcttta ccaatgacac tttggaaagt tctaaccaaa gttggatatg ggtgaggacc
2941 aatggcagaa ccgacgacgt agtaagtagt tttcaagtta gtaacccaaa atctgaatgc
3001 ctctgaagta gcgtctctta gagtctttgt accattagta acagcaatta ctttagcacc
3061 gagaattctc attctgaaga cgtttaaagc ttggcgacga acatcttctg cacccatgaa
3121 cacagtacag gttaagccaa atttagcaca tgcagtggca gtggcaacac cgtgttgacc
3181 agcaccggtt tcagcaataa cgttcttctt gcctaatctt ttagctagaa gaacttgtgc
3241 taaagcattg ttgatctagt gagatcccgt gtggttaaga tcttctctct tcaaccagat
3301 ttgagcacct tgacaatgct cagttaatct ctcagctttg tgtagtgaag aaggacggcc
3361 aatataagaa tacaaggatt tgaagtcttc ccagaatgtg ggatcggcga cagcttcatc
3421 aaaacccttt tccaactctc ttagacatgc atgaagagct tctgggacat actgaccacc
3481 aaagtcccca aatctaattg ggtgcttgtg attttcatca aattcgtcca gtatttcctt
3541 cttaacgttt gcggacttca aggactcttt ttgaaaggca aagaattcgt ccttggacag
3601 aaccttatgc ttagcaccat ttagaattcc ctgtacatat tccttagcaa cgtcgtacct
3661 tttgccctct ggagcatctc cacataatgt gacgattttg gaaccaatca ctacaccgtc
3721 agcaacacta ccaactgatt ggaaatgttc tctggtagag acaccaaacc caacggccaa
3781 aggagtatcc ttggtgtact ttctgactct agagatgagt tcatccaaat cactggccac
3841 agaactttga acaccagtag tacccattct agacacaacg tagacaaacg aatcggcaat
3901 atgcgatagt aattccaatc tttcgtcggt ggtagaagga gccactagtg ggatcaggct
3961 caaaccatta tcattgatgt agtttctgac cttcaacgcc tcctctggtg gcaaatcgac
4021 gatgataaaa ccattagcac cagccttggc agcgtcctga ataaatcttt cttcaccgta
4081 gtttagaata gggttatagt aacccattag gattatgggt acggtaacac cttcatttct
4141 agcttgggag accatttcta gagtttgagg caaggtaaca ccgttttgca aagccacagt
4201 attagataat tgaattgtag gaccatctgc aattggatca gagaagggca tacccaattc
4261 gatgatatct acaccaccat cctggaaacc cttgagaata gggacagtgt ctttgactgt
4321 tgggtaacct gcggtcataa atgtgaccaa ggcgttcctg ttttcttttt tagcgttagc
4381 aaatgtttgt ctgagttgtt ctgacatttt acatgtctat gagcttatca catgtttcta
4441 catttttctg atatttgaaa gttattaatc tttttgtttc tatagatctt cctatgtata
4501 ggtcatccct cagactcaga ttggtgtttc aaagatcctc aaggcaatag aataacagat
4561 tttgatagta ttaataaaga attaggtctt ggtagaagag atgtaaaatt agataaaggt
4621 catgatgatt taattaaatt atgtactgaa aaaatagata gtactattaa ttgcgttgcg
4681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
4741 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
4801 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
4861 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
4921 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
4981 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
5041 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
5101 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
5161 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
5221 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
5281 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
5341 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
5401 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
5461 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
5521 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
5581 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
5641 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
5701 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
5761 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
5821 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
5881 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
5941 atccacctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
6001 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
6061 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
6121 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
6181 cgcagtgtta tcactcatgg ttatggcagc actgcataat
//
