| 质粒名称: | p112A1NE、p112AINE |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 85983 |
| 存储实验室: | Wolf Frommer |
|
载体骨架基本信息 |
|
| 载体名称: | p112A1NE |
| 载体类型: | 酵母表达 |
| 筛选标记: | TRP1 |
| 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 |
|
| 启动子: | Adh1 |
| 克隆方法: | 限制性内切酶 |
|
文献引用方法 |
|
| 材料和方法部分: | p112A1NE was a gift from Wolf Frommer (Addgene plasmid # 85983 ; http://n2t.net/addgene:85983 ; RRID:Addgene_85983) |
| 参考文献部分: | Isolation and characterization of a sucrose carrier cDNA from spinach by functional expression in yeast. Riesmeier JW, Willmitzer L, Frommer WB. EMBO J. 1992 Dec;11(13):4705-13. PubMed 1464305 |
买家导航
p112AINE载体基本信息
p112AINE载体图谱质粒图谱和多克隆位点信息
p112AINE载体简介
p112AINE载体序列
LOCUS p112A1NE 5725 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION For construction of cDNA expression library..
ACCESSION .
VERSION .
KEYWORDS p112A1NE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5725)
AUTHORS Riesmeier JW, Willmitzer L, Frommer WB
TITLE Isolation and characterization of a sucrose carrier cDNA from
spinach by functional expression in yeast.
JOURNAL EMBO J. 1992 Dec;11(13):4705-13.
PUBMED 1464305
REFERENCE 2 (bases 1 to 5725)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5725
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 113..132
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 366..383
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 500..521
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 536..566
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 574..590
/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)."
primer_bind 579..601
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 598..614
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 598..614
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 638..1034
/gene="S. cerevisiae ADH1"
/label=ADH1 promoter
/note="promoter for alcohol dehydrogenase 1"
terminator 1221..1408
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(1276..1300)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
primer_bind complement(1420..1437)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1420..1436)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1429..1451)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 1873..3219
/label=2u ori
/note="yeast 2u plasmid origin of replication"
CDS complement(3331..4005)
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
promoter complement(4006..4107)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
primer_bind complement(4127..4145)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4213..4317
/gene="bla"
/label=AmpR promoter
CDS 4318..5178
/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 complement(4536..4555)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5349..212
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
61 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
181 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
241 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
301 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
361 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
421 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
481 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
541 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
601 gaaacagcta tgaccatgat tacgccaagc ttgcatgcaa cttcttttct ttttttttct
661 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
721 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
781 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
841 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
901 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
961 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
1021 gcatacaatc aactccaagc ttcagctgct gcagtctaga gaattcccgg gcggccgcgg
1081 atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct
1141 tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg
1201 ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
1261 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
1321 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
1381 tcgctcttat tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg
1441 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
1501 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
1561 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
1621 acaccgcata tatcggatcg tacttgttac ccatcattga attttgaaca tccgaacctg
1681 ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat agtctagcgc
1741 tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc tattgcatag
1801 gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc acttcaatag
1861 catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
1921 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
1981 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
2041 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
2101 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
2161 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
2221 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
2281 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
2341 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
2401 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
2461 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
2521 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
2581 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
2641 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
2701 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
2761 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt
2821 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcggaatag
2881 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
2941 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
3001 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
3061 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
3121 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
3181 ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat aagctgtcaa
3241 acatgagaat tgatctttta tgcttgcttt tcaaaaggct tgcaggcaag tgcacaaaca
3301 atacttaaat aaatactact cagtaataac ctatttctta gcatttttga cgaaatttgc
3361 tattttgtta gagtctttta caccatttgt ctccacacct ccgcttacat caacaccaat
3421 aacgccattt aatctaagcg catcaccaac attttctggc gtcagtccac cagctaacat
3481 aaaatgtaag ctctcggggc tctcttgcct tccaacccag tcagaaatcg agttccaatc
3541 caaaagttca cctgtcccac ctgcttctga atcaaacaag ggaataaacg aatgaggttt
3601 ctgtgaagct gcactgagta gtatgttgca gtcttttgga aatacgagtc ttttaataac
3661 tggcaaaccg aggaactctt ggtattcttg ccacgactca tctccatgca gttggacgat
3721 atcaatgccg taatcattga ccagagccaa aacatcctcc ttaggttgat tacgaaacac
3781 gccaaccaag tatttcggag tgcctgaact atttttatat gcttttacaa gacttgaaat
3841 tttccttgca ataaccgggt caattgttct ctttctattg ggcacacata taatacccag
3901 caagtcagca tcggaatcta gtgcacattc tgcggcctct gtgctctgca agccgcaaac
3961 tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc aagctgcctt
4021 tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc cctcttggcc
4081 ctctcctttt cttttttcga ccgaattaat tcttgaagac gaaagggcct cgtgatacgc
4141 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
4201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
4261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
4321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
4381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
4441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
4501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
4561 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
4621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
4681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
4741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
4801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
4861 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
4921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
4981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
5041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
5101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
5161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
5221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
5281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
5341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
5401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
5461 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
5521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
5581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
5641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
5701 cgaacgacct acaccgaact gagat
//
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