| 质粒名称: | pETcon(-) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 41522 |
| 存储实验室: | Andrew Scharenberg |
|
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: | 6139 bp |
| 完整质粒大小: | -- |
| 载体修饰: | Stuffer inserted between NdeI and XhoI (puts Myc tag out of frame to prevent parental contamination). Originally derived from the pCTcon2 vector. |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | GAL1 |
| 标签/融合蛋白: |
HA（骨干网上的N个终端） 3x(G4S) Linker（主干上的N个终端） Stuffer （插入件上的N端子） G3S Linker （主干上的C终端） Myc Tag（主干网上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | GAL1 |
| 3' 测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pETcon(-) was a gift from Andrew Scharenberg (Addgene plasmid # 41522 ; http://n2t.net/addgene:41522 ; RRID:Addgene_41522) |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pETcon(-) 6173 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pETcon(-)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6173)
TITLE pETCON Yeast Surface Display Plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6173)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6173
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 2..18
/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 7..29
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 26..42
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 26..42
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 63..81
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 315..756
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 315..432
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 727..750
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
CDS 1082..1093
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 1094..1120
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1236..1265
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(1594..1613)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1595..1613)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1620..1637)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1620..1636)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1629..1651)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 1777..2232
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1864..1883)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2074..2095
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(2335..3009)
/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(3010..3290)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
primer_bind complement(3311..3330)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3430..3452
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3490..3508)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 3549..4052
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 4089..4193
/gene="bla"
/label=AmpR promoter
CDS 4194..5054
/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(4412..4431)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5225..5813
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5714..5733
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5967..5984
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6101..6122
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6137..6167
/label=lac promoter
/note="promoter for the E. coli lac operon"
ORIGIN
1 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
61 gaaattaacc ctcactaaag ggaacaaaag ctggtaccaa ttccttgaat tttcaaaaat
121 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
181 accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
241 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
301 gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
361 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
421 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
481 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
541 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
601 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
661 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
721 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat cgaattccct
781 acttcataca ttttcaatta agatgcagtt acttcgctgt ttttcaatat tttctgttat
841 tgcttcagtt ttagcacagg aactgacaac tatatgcgag caaatcccct caccaacttt
901 agaatcgacg ccgtactctt tgtcaacgac tactattttg gccaacggga aggcaatgca
961 aggagttttt gaatattaca aatcagtaac gtttgtcagt aattgcggtt ctcacccctc
1021 aacaactagc aaaggcagcc ccataaacac acagtatgtt tttaaggaca atagctcgac
1081 gattgaaggt agatacccat acgacgttcc agactacgct ctgcaggcta gtggtggagg
1141 aggctctggt ggaggcggta gcggaggcgg agggtcggct agccatatgt gaagcatgct
1201 taattaaggc gccttatctc gaggggggcg gatccgaaca aaagcttatt tctgaagagg
1261 acttgtaata gagatctgat aacaacagtg tagatgtaac aaaatcgact ttgttcccac
1321 tgtactttta gctcgtacaa aatacaatat acttttcatt tctccgtaaa caacatgttt
1381 tcccatgtaa tatccttttc tatttttcgt tccgttacca actttacaca tactttatat
1441 agctattcac ttctatacac taaaaaacta agacaatttt aattttgctg cctgccatat
1501 ttcaatttgt tataaattcc tataatttat cctattagta gctaaaaaaa gatgaatgtg
1561 aatcgaatcc taagagaatt gagctccaat tcgccctata gtgagtcgta ttacaattca
1621 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
1681 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
1741 ctttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
1801 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
1861 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
1921 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
1981 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
2041 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
2101 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
2161 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
2221 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
2281 catagatcgg caagtgcaca aacaatactt aaataaatac tactcagtaa taacctattt
2341 cttagcattt ttgacgaaat ttgctatttt gttagagtct tttacaccat ttgtctccac
2401 acctccgctt acatcaacac caataacgcc atttaatcta agcgcatcac caacattttc
2461 tggcgtcagt ccaccagcta acataaaatg taagctttcg gggctctctt gccttccaac
2521 ccagtcagaa atcgagttcc aatccaaaag ttcacctgtc ccacctgctt ctgaatcaaa
2581 caagggaata aacgaatgag gtttctgtga agctgcactg agtagtatgt tgcagtcttt
2641 tggaaatacg agtcttttaa taactggcaa accgaggaac tcttggtatt cttgccacga
2701 ctcatctcca tgcagttgga cgatatcaat gccgtaatca ttgaccagag ccaaaacatc
2761 ctccttaggt tgattacgaa acacgccaac caagtatttc ggagtgcctg aactattttt
2821 atatgctttt acaagacttg aaattttcct tgcaataacc gggtcaattg ttctctttct
2881 attgggcaca catataatac ccagcaagtc agcatcggaa tctagagcac attctgcggc
2941 ctctgtgctc tgcaagccgc aaactttcac caatggacca gaactacctg tgaaattaat
3001 aacagacata ctccaagctg cctttgtgtg cttaatcacg tatactcacg tgctcaatag
3061 tcaccaatgc cctccctctt ggccctctcc ttttcttttt tcgaccgaat taattcttaa
3121 tcggcaaaaa aagaaaagct ccggatcaag attgtacgta aggtgacaag ctatttttca
3181 ataaagaata tcttccacta ctgccatctg gcgtcataac tgcaaagtac acatatatta
3241 cgatgctgtt ctattaaatg cttcctatat tatatatata gtaatgtcgt gatctatggt
3301 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
3361 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
3421 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
3481 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
3541 cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa
3601 tttgggaatt tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta
3661 aataaagaag gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa
3721 tttcaacaaa aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag
3781 atatacattc gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt
3841 ctacacagac aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa
3901 aaggtagtat ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta
3961 ttttttcttt aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa
4021 tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc acttttcggg
4081 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
4141 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
4201 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
4261 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
4321 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
4381 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
4441 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
4501 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
4561 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
4621 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
4681 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
4741 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
4801 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
4861 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
4921 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
4981 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
5041 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
5101 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
5161 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
5221 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
5281 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
5341 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
5401 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
5461 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
5521 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
5581 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
5641 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
5701 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
5761 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
5821 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
5881 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
5941 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
6001 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
6061 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
6121 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt gga
//
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