| 质粒名称: | pETCON-HtsptLB12v.3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 45121 |
| 存储实验室: | David Baker |
|
载体骨架基本信息 |
|
| 载体名称: | pETCON |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6287 bp |
| 完整质粒大小: | 6767 bp |
| 载体修饰: | -- |
| 载体类型: | 酵母表达 |
| 筛选标记: | TRP1 |
| 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: | HtsptLB12v.3 |
| 物种来源: | Synthetic |
| 插入大小: | 480 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | GAL |
| 标签/融合蛋白: |
c-myc epitope tag（主干上的c端）
Aga2p for yeast surface display（主干上的N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NdeI(未破坏) |
| 3' 克隆位点: | XhoI(未破坏) |
| 5' 测序引物 | -- |
| 3' 测序引物 | cgagctaaaagtacagtggg |
|
文献引用方法 |
|
| 材料和方法部分: | pETCON-HtsptLB12v.3 was a gift from David Baker (Addgene plasmid # 45121 ; http://n2t.net/addgene:45121 ; RRID:Addgene_45121) |
买家导航
pETCONHtsptLB12v.3载体基本信息
pETCONHtsptLB12v.3载体图谱质粒图谱和多克隆位点信息
pETCONHtsptLB12v.3载体简介
pETCONHtsptLB12v.3载体序列
LOCUS pETCON-HtsptLB12v.3 6767 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pETCON-HtsptLB12v.3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6767)
TITLE Expression plasmids for designed HEL-binding proteins
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6767)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6767
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..28)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 70..573
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 610..714
/gene="bla"
/label=AmpR promoter
CDS 715..1575
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(933..952)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1746..2334
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2488..2505
/label=L4440
/note="L4440 vector, forward primer"
promoter 2658..2688
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2696..2712
/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 2701..2723
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2720..2736
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2720..2736
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 2757..2775
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 2940..3604
/gene="GAL1 and GAL10 genes of S. cerevisiae"
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
protein_bind 3156..3273
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 3568..3591
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
CDS 3923..3934
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 3935..3961
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 4529..4558
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(4887..4906)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4888..4906)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4913..4930)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4913..4929)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4922..4944)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5074..5529
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5161..5180)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5371..5392
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(5626..6300)
/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(6301..6581)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
primer_bind complement(6598..6617)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6717..6739
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt
1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
2101 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
2221 agggagcttc caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc
2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaatt
2761 aaccctcact aaagggaaca aaagctgggt acccgacagg ttatcagcaa caacacagtc
2821 atatccattc tcaattagct ctaccacagt gtgtgaacca atgtatccag caccacctgt
2881 aaccaaaaca attttagaag tactttcact ttgtaactga gctgtcattt atattgaatt
2941 ttcaaaaatt cttacttttt ttttggatgg acgcaaagaa gtttaataat catattacat
3001 ggcattacca ccatatacat atccatatac atatccatat ctaatcttac ttatatgttg
3061 tggaaatgta aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag
3121 taatacgctt aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggtg
3181 acagccctcc gaaggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg
3241 aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct
3301 tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaatgaac
3361 gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg
3421 taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat gcaaaaactg
3481 cataaccact ttaactaata ctttcaacat tttcggtttg tattacttct tattcaaatg
3541 taataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa
3601 aaaccccgga tcgaattccc tacttcatac attttcaatt aagatgcagt tacttcgctg
3661 tttttcaata ttttctgtta ttgcttcagt tttagcacag gaactgacaa ctatatgcga
3721 gcaaatcccc tcaccaactt tagaatcgac gccgtactct ttgtcaacga ctactatttt
3781 ggccaacggg aaggcaatgc aaggagtttt tgaatattac aaatcagtaa cgtttgtcag
3841 taattgcggt tctcacccct caacaactag caaaggcagc cccataaaca cacagtatgt
3901 ttttaaggac aatagctcga cgattgaagg tagataccca tacgacgttc cagactacgc
3961 tctgcaggct agtggtggag gaggctctgg tggaggcggt agcggaggcg gagggtcggc
4021 tagccatatg ggttccgaaa tgagtgaaat gtctgttgta gattacgtcg ttaagtcttt
4081 taattataca gaggatatga gtcatgaatt gttattctct gttaaaaaga gatggttcgt
4141 aaagccattc gaacactcaa ccagatatgg tttgttagcc tacatgttgt tacctggtaa
4201 ctacataaag ttcggtttgt acgttttaac gaaccaagac tatgctagat tcgaaatagc
4261 atgggtacat gtcgataaag acggtaaaat cgaagaaaga accgtatatt ccattgaaac
4321 ttactggcac attttcattg atatcgaaaa cgacttgaac tgtccatacg ttttggcaaa
4381 gtttattgaa atgagacctg aattcgataa gacagcctgg gtcgaagaat caaactattc
4441 cattgctgaa gatgacatcc aaatggttga aagtataaaa agatacttgg aaagaaagat
4501 cgcttctgat ctcgaggggg gcggatccga acaaaagctt atttctgaag aggacttgta
4561 atagagatct gataacaaca gtgtagatgt aacaaaatcg actttgttcc cactgtactt
4621 ttagctcgta caaaatacaa tatacttttc atttctccgt aaacaacatg ttttcccatg
4681 taatatcctt ttctattttt cgttccgtta ccaactttac acatacttta tatagctatt
4741 cacttctata cactaaaaaa ctaagacaat tttaattttg ctgcctgcca tatttcaatt
4801 tgttataaat tcctataatt tatcctatta gtagctaaaa aaagatgaat gtgaatcgaa
4861 tcctaagaga attgagctcc aattcgccct atagtgagtc gtattacaat tcactggccg
4921 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
4981 cacatccccc cttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
5041 aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg
5101 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
5161 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
5221 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
5281 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
5341 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
5401 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
5461 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
5521 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcag
5581 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
5641 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
5701 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
5761 tccaccagct aacataaaat gtaagctttc ggggctctct tgccttccaa cccagtcaga
5821 aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat
5881 aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac
5941 gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc
6001 atgcagttgg acgatatcaa tgccgtaatc attgaccaga gccaaaacat cctccttagg
6061 ttgattacga aacacgccaa ccaagtattt cggagtgcct gaactatttt tatatgcttt
6121 tacaagactt gaaattttcc ttgcaataac cgggtcaatt gttctctttc tattgggcac
6181 acatataata cccagcaagt cagcatcgga atctagagca cattctgcgg cctctgtgct
6241 ctgcaagccg caaactttca ccaatggacc agaactacct gtgaaattaa taacagacat
6301 actccaagct gcctttgtgt gcttaatcac gtatactcac gtgctcaata gtcaccaatg
6361 ccctccctct tggccctctc cttttctttt ttcgaccgaa ttaattctta atcggcaaaa
6421 aaagaaaagc tccggatcaa gattgtacgt aaggtgacaa gctatttttc aataaagaat
6481 atcttccact actgccatct ggcgtcataa ctgcaaagta cacatatatt acgatgctgt
6541 ctattaaatg cttcctatat tatatatata gtaatgtcgt ttatggtgca ctctcagtac
6601 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
6661 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
6721 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
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