| 质粒名称: | pCT302 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 41845 |
| 存储实验室: | Dane Wittrup |
|
载体骨架基本信息 |
|
| 载体名称: | pCT |
| 载体出品公司: | -- |
| 空载体大小: | 6142 bp |
| 完整质粒大小: | 6970 bp |
| 载体修饰: | -- |
| 载体类型: | 酵母表达 |
| 筛选标记: | TRP1 |
| 载体抗性: | 氨苄霉素,100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | 4-4-20 |
| 物种来源: | Synthetic |
| 插入大小: | 765 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: |
HA（主干上的N终端）
c-myc（主干上的c终端）
|
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCT302 was a gift from Dane Wittrup (Addgene plasmid # 41845 ; http://n2t.net/addgene:41845 ; RRID:Addgene_41845) |
| 参考文献部分: | Substantial energetic improvement with minimal structural perturbation in a high affinity mutant antibody. Midelfort KS, Hernandez HH, Lippow SM, Tidor B, Drennan CL, Wittrup KD. J Mol Biol. 2004 Oct 22;343(3):685-701. 10.1016/j.jmb.2004.08.019 PubMed 15465055 |
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 pCT302 7032 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCT302
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7032)
AUTHORS Midelfort KS, Hernandez HH, Lippow SM, Tidor B, Drennan CL, Wittrup
KD
TITLE Substantial energetic improvement with minimal structural
perturbation in a high affinity mutant antibody.
JOURNAL J Mol Biol. 2004 Oct 22;343(3):685-701.
PUBMED 15465055
REFERENCE 2 (bases 1 to 7032)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7032
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(159..833)
/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(834..1114)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
primer_bind complement(1131..1150)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1250..1272
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1310..1328)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 1370..1873
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 1910..2014
/gene="bla"
/label=AmpR promoter
CDS 2015..2875
/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(2233..2252)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3046..3634
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3788..3805
/label=L4440
/note="L4440 vector, forward primer"
promoter 3958..3988
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3996..4012
/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 4001..4023
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4020..4036
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4020..4036
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 4057..4075
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 4240..4904
/gene="GAL1 and GAL10 genes of S. cerevisiae"
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
protein_bind 4456..4573
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 4868..4891
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
CDS 5223..5234
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 5235..5261
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 6090..6119
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(6452..6471)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6453..6471)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6478..6495)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6478..6494)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6487..6509)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 6639..62
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6726..6745)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6936..6957
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
61 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caggcaagtg
121 cacaaacaat acttaaataa atactactca gtaataacct atttcttagc atttttgacg
181 aaatttgcta ttttgttaga gtcttttaca ccatttgtct ccacacctcc gcttacatca
241 acaccaataa cgccatttaa tctaagcgca tcaccaacat tttctggcgt cagtccacca
301 gctaacataa aatgtaagct ttcggggctc tcttgccttc caacccagtc agaaatcgag
361 ttccaatcca aaagttcacc tgtcccacct gcttctgaat caaacaaggg aataaacgaa
421 tgaggtttct gtgaagctgc actgagtagt atgttgcagt cttttggaaa tacgagtctt
481 ttaataactg gcaaaccgag gaactcttgg tattcttgcc acgactcatc tccatgcagt
541 tggacgatat caatgccgta atcattgacc agagccaaaa catcctcctt aggttgatta
601 cgaaacacgc caaccaagta tttcggagtg cctgaactat ttttatatgc ttttacaaga
661 cttgaaattt tccttgcaat aaccgggtca attgttctct ttctattggg cacacatata
721 atacccagca agtcagcatc ggaatctaga gcacattctg cggcctctgt gctctgcaag
781 ccgcaaactt tcaccaatgg accagaacta cctgtgaaat taataacaga catactccaa
841 gctgcctttg tgtgcttaat cacgtatact cacgtgctca atagtcacca atgccctccc
901 tcttggccct ctccttttct tttttcgacc gaattaattc ttaatcggca aaaaaagaaa
961 agctccggat caagattgta cgtaaggtga caagctattt ttcaataaag aatatcttcc
1021 actactgcca tctggcgtca taactgcaaa gtacacatat attacgatgc tgtctattaa
1081 atgcttccta tattatatat atagtaatgt cgtttatggt gcactctcag tacaatctgc
1141 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
1201 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
1261 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
1321 cgcctatttt tataggttaa tgtcatgata ataatggttt cttaggacgg atcgcttgcc
1381 tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg
1441 tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag
1501 ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact
1561 ttacatatat atttattaga caagaaaagc agattaaata gatatacatt cgattaacga
1621 taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa
1681 caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg
1741 atccccctag agtcttttac atcttcggaa aacaaaaact attttttctt taatttcttt
1801 ttttactttc tatttttaat ttatatattt atattaaaaa atttaaatta taattatttt
1861 tatagcacgt gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
1921 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
1981 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
2041 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
2101 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
2161 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
2221 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
2281 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
2341 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
2401 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
2461 ttttcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
2521 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
2581 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
2641 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
2701 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
2761 agatggtaag ccctcccgta tcgtagttat ctacacgacg ggcagtcagg caactatgga
2821 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
2881 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
2941 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
3001 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
3061 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
3121 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
3181 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
3241 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
3301 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
3361 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
3421 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
3481 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggggaa
3541 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
3601 gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc agcaacgcgg cctttttacg
3661 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
3721 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
3781 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
3841 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
3901 gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac cccaggcttt
3961 acactttatg cttccggctc ctatgttgtg tggaattgtg agcggataac aatttcacac
4021 aggaaacagc tatgaccatg attacgccaa gctcggaatt aaccctcact aaagggaaca
4081 aaagctgggt acccgacagg ttatcagcaa caacacagtc atatccattc tcaattagct
4141 ctaccacagt gtgtgaacca atgtatccag caccacctgt aaccaaaaca attttagaag
4201 tactttcact ttgtaactga gctgtcattt atattgaatt ttcaaaaatt cttacttttt
4261 ttttggatgg acgcaaagaa gtttaataat catattacat ggcattacca ccatatacat
4321 atccatatac atatccatat ctaatcttac ttatatgttg tggaaatgta aagagcccca
4381 ttatcttagc ctaaaaaaac cttctctttg gaactttcag taatacgctt aactgctcat
4441 tgctatattg aagtacggat tagaagccgc cgagcgggtg acagccctcc gaaggaagac
4501 tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg
4561 ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta tgaagaggaa
4621 aaattggcag taacctggcc ccacaaacct tcaaatgaac gaatcaaatt aacaaccata
4681 ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca gcgaagcgat
4741 gatttttgat ctattaacag atatataaat gcaaaaactg cataaccact ttaactaata
4801 ctttcaacat tttcggtttg tattacttct tattcaaatg taataaaagt atcaacaaaa
4861 aattgttaat atacctctat actttaacgt caaggagaaa aaaccccgga tcgaattccc
4921 tacttcatac attttcaatt aagatgcagt tacttcgctg tttttcaata ttttctgtta
4981 ttgcttcagt tttagcacag gaactgacaa ctatatgcga gcaaatcccc tcaccaactt
5041 tagaatcgac gccgtactct ttgtcaacga ctactatttt ggccaacggg aaggcaatgc
5101 aaggagtttt tgaatattac aaatcagtaa cgtttgtcag taattgcggt tctcacccct
5161 caacaactag caaaggcagc cccataaaca cacagtatgt ttttaaggac aatagctcga
5221 cgattgaagg tagataccca tacgacgttc cagactacgc tctgcaggct agtggtggtg
5281 gtggttctgg tggtggtggt tctggtggtg gtggttctgc tagcgacgtc gttatgactc
5341 aaacaccact atcacttcct gttagtctag gtgatcaagc ctccatctct tgcagatcta
5401 gtcagagcct tgtacacagt aatggaaaca cctatttacg ttggtacctg cagaagccag
5461 gccagtctcc aaaggtcctg atctacaaag tttccaaccg attttctggg gtcccagaca
5521 ggttcagtgg cagtggatca gggacagatt tcacactcaa gatcagcaga gtggaggctg
5581 aggatctggg agtttatttc tgctctcaaa gtacacatgt tccgtggacg ttcggtggag
5641 gcaccaagct tgaaattaag tcctctgctg atgatgctaa gaaggatgct gctaagaagg
5701 atgatgctaa gaaagatgat gctaagaaag atggtgacgt caaactggat gagactggag
5761 gaggcttggt gcaacctggg aggcccatga aactctcctg tgttgcctct ggattcactt
5821 ttagtgacta ctggatgaac tgggtccgcc agtctccaga gaaaggactg gagtgggtag
5881 cacaaattag aaacaaacct tataattatg aaacatatta ttcagattct gtgaaaggca
5941 gattcaccat ctcaagagat gattccaaaa gtagtgtcta cctgcaaatg aacaacttaa
6001 gagttgaaga catgggtatc tattactgta cgggttctta ctatggtatg gactactggg
6061 gtcaaggaac ctcagtcacc gtctcctcag aacaaaagct tatttctgaa gaagacttgt
6121 aatagctcga gatctgataa caacagtgta gatgtaacaa aatcgacttt gttcccactg
6181 tacttttagc tcgtacaaaa tacaatatac ttttcatttc tccgtaaaca acatgttttc
6241 ccatgtaata tccttttcta tttttcgttc cgttaccaac tttacacata ctttatatag
6301 ctattcactt ctatacacta aaaaactaag acaattttaa ttttgctgcc tgccatattt
6361 caatttgtta taaattccta taatttatcc tattagtagc taaaaaaaga tgaatgtgaa
6421 tcgaatccta agagaattga gctccaattc gccctatagt gagtcgtatt acaattcact
6481 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
6541 tgcagcacat ccccccttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
6601 ttcccaacag ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta gcggcgcatt
6661 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
6721 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
6781 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
6841 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
6901 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
6961 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
7021 ctattggtta aa
//
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