| 质粒名称: | p406TEF1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 15976 |
| 存储实验室: | Nicolas Buchler, Fred Cross |
|
载体骨架基本信息 |
|
| 载体名称: | pRS406 |
| 载体出品公司: | Sikorski & Hieter, Genetics 122:19-27 (1989) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4283 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，酵母表达 |
| 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: | TEF1 promoter + polylinker + CYC1 terminator,Translational elongation factor EF-1 alpha |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 728 bp |
| 基因突变: | TEF1 promoter is between SacI and XbaI restriction sites. |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SacI(未破坏) |
| 3' 克隆位点: | KpnI(未破坏) |
| 5' 测序引物 | M13 Reverse |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | p406TEF1 was a gift from Nicolas Buchler & Fred Cross (Addgene plasmid # 15976 ; http://n2t.net/addgene:15976 ; RRID:Addgene_15976) |
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 p406TEF1 5010 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS p406TEF1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5010)
TITLE Buchler lab plasmids
REFERENCE 2 (bases 1 to 5010)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5010
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..28)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 96..200
/gene="bla"
/label=AmpR promoter
CDS 201..1061
/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(419..438)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1232..1820
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1721..1740
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1974..1991
/label=L4440
/note="L4440 vector, forward primer"
promoter 2144..2174
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2182..2198
/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 2187..2209
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2206..2222
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2206..2222
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2241..2261
/label=T3
/note="T3 promoter, forward primer"
promoter 2243..2261
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2279..2677
/gene="S. cerevisiae TEF1"
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
primer_bind 2681..2697
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2681..2697
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(2731..2747)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2732..2748)
/label=pBluescriptKS
/note="For pBluescript vector"
terminator 2750..2997
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(2766..2784)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind complement(3015..3034)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3016..3034)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3044..3061)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3044..3060)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3053..3075)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3205..3660
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3292..3311)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3502..3523
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(3791..4594)
/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="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(4595..4810)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
primer_bind complement(4841..4860)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4960..4982
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
2101 acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctca
2281 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
2341 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
2401 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
2461 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
2521 ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt
2581 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
2641 tgctcattag aaagaaagca tagcaatcta atctaagttt tctagaacta gtggatcccc
2701 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagt catgtaatta
2761 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
2821 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
2881 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
2941 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc
3001 cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
3061 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
3121 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
3181 gcagcctgaa tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
3241 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
3301 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
3361 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
3421 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
3481 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
3541 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
3601 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
3661 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagggtaata
3721 actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata
3781 atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc
3841 tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa
3901 taataatgtc agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg
3961 cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat
4021 ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg
4081 caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt
4141 ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac
4201 cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta
4261 ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc
4321 tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc
4381 cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac
4441 ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca
4501 agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag
4561 tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt
4621 ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata
4681 tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga
4741 ttaccgaatc aaaaaaattt caaagaaacc gaaatcaaaa aaaagaataa aaaaaaaatg
4801 atgaattgaa ttgaaaagct gtggtatggt gcactctcag tacaatctgc tctgatgccg
4861 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
4921 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
4981 ggttttcacc gtcatcaccg aaacgcgcga
//
