| 质粒名称: | pCTcon2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 41843 |
| 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: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6142 bp |
| 完整质粒大小: | 6456 bp |
| 载体修饰: | -- |
| 载体类型: | 酵母表达 |
| 筛选标记: | TRP1 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: |
CD20
|
| 物种来源: | -- |
| 插入大小: | 183 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: | pCTcon2 was a gift from Dane Wittrup (Addgene plasmid # 41843 ; http://n2t.net/addgene:41843 ; RRID:Addgene_41843) |
| 参考文献部分: | Isolating and engineering human antibodies using yeast surface display. Chao G, Lau WL, Hackel BJ, Sazinsky SL, Lippow SM, Wittrup KD. Nat Protoc. 2006;1(2):755-68. 10.1038/nprot.2006.94 PubMed 17406305 |
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 pCTcon2 6314 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCTcon2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6314)
AUTHORS Chao G, Lau WL, Hackel BJ, Sazinsky SL, Lippow SM, Wittrup KD
TITLE Isolating and engineering human antibodies using yeast surface
display.
JOURNAL Nat Protoc. 2006;1(2):755-68.
PUBMED 17406305
REFERENCE 2 (bases 1 to 6314)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6314
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..442
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 1..118
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 413..436
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
CDS 768..779
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 780..806
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1059..1088
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(1421..1440)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1422..1440)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1447..1464)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1447..1463)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1456..1478)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 1604..2059
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1691..1710)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1901..1922
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(2162..2836)
/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(2837..3117)
/gene="S. cerevisiae TRP1"
/label=TRP1 promoter
primer_bind complement(3138..3157)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3257..3279
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3317..3335)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
misc_feature 3376..3879
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 3916..4020
/gene="bla"
/label=AmpR promoter
CDS 4021..4881
/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(4239..4258)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5052..5640
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5541..5560
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5794..5811
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5928..5949
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5964..5994
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6002..6018
/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 6007..6029
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6026..6042
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6026..6042
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 6063..6081
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
421 tctatacttt aacgtcaagg agaaaaaacc ccggatcgaa ttccctactt catacatttt
481 caattaagat gcagttactt cgctgttttt caatattttc tgttattgct tcagttttag
541 cacaggaact gacaactata tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt
601 actctttgtc aacgactact attttggcca acgggaaggc aatgcaagga gtttttgaat
661 attacaaatc agtaacgttt gtcagtaatt gcggttctca cccctcaaca actagcaaag
721 gcagccccat aaacacacag tatgttttta aggacaatag ctcgacgatt gaaggtagat
781 acccatacga cgttccagac tacgctctgc aggctagtgg tggaggaggc tctggtggag
841 gcggtagcgg aggcggaggg tcggctagct gcggtggcgg cggtactagc aaaatttctc
901 attttttgaa aatggaatct ttgaatttta ttagagctca tactccatat attaatattt
961 acaattgtga accagctaat ccatctgaaa aaaattctcc atctactcaa tattgttatt
1021 ctattcaatc ttcccaggtc gactgcgggg gcggatccga acaaaagctt atttctgaag
1081 aggacttgta atagctcgag atctgataac aacagtgtag atgtaacaaa atcgactttg
1141 ttcccactgt acttttagct cgtacaaaat acaatatact tttcatttct ccgtaaacaa
1201 catgttttcc catgtaatat ccttttctat ttttcgttcc gttaccaact ttacacatac
1261 tttatatagc tattcacttc tatacactaa aaaactaaga caattttaat tttgctgcct
1321 gccatatttc aatttgttat aaattcctat aatttatcct attagtagct aaaaaaagat
1381 gaatgtgaat cgaatcctaa gagaattgag ctccaattcg ccctatagtg agtcgtatta
1441 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
1501 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
1561 cgatcgcctt tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
1621 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
1681 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
1741 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
1801 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
1861 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
1921 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
1981 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
2041 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
2101 cacaccgcat agatcggcaa gtgcacaaac aatacttaaa taaatactac tcagtaataa
2161 cctatttctt agcatttttg acgaaatttg ctattttgtt agagtctttt acaccatttg
2221 tctccacacc tccgcttaca tcaacaccaa taacgccatt taatctaagc gcatcaccaa
2281 cattttctgg cgtcagtcca ccagctaaca taaaatgtaa gctttcgggg ctctcttgcc
2341 ttccaaccca gtcagaaatc gagttccaat ccaaaagttc acctgtccca cctgcttctg
2401 aatcaaacaa gggaataaac gaatgaggtt tctgtgaagc tgcactgagt agtatgttgc
2461 agtcttttgg aaatacgagt cttttaataa ctggcaaacc gaggaactct tggtattctt
2521 gccacgactc atctccatgc agttggacga tatcaatgcc gtaatcattg accagagcca
2581 aaacatcctc cttaggttga ttacgaaaca cgccaaccaa gtatttcgga gtgcctgaac
2641 tatttttata tgcttttaca agacttgaaa ttttccttgc aataaccggg tcaattgttc
2701 tctttctatt gggcacacat ataataccca gcaagtcagc atcggaatct agagcacatt
2761 ctgcggcctc tgtgctctgc aagccgcaaa ctttcaccaa tggaccagaa ctacctgtga
2821 aattaataac agacatactc caagctgcct ttgtgtgctt aatcacgtat actcacgtgc
2881 tcaatagtca ccaatgccct ccctcttggc cctctccttt tcttttttcg accgaattaa
2941 ttcttaatcg gcaaaaaaag aaaagctccg gatcaagatt gtacgtaagg tgacaagcta
3001 tttttcaata aagaatatct tccactactg ccatctggcg tcataactgc aaagtacaca
3061 tatattacga tgctgttcta ttaaatgctt cctatattat atatatagta atgtcgtgat
3121 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
3181 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
3241 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
3301 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
3361 atggtttctt agacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat
3421 ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta
3481 gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt
3541 taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat
3601 taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg
3661 tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc
3721 aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca
3781 aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat atatttatat
3841 taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact
3901 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
3961 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
4021 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
4081 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
4141 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
4201 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
4261 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
4321 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
4381 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
4441 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
4501 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
4561 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
4621 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
4681 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
4741 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
4801 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
4861 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
4921 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
4981 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
5041 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
5101 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
5161 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
5221 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
5281 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
5341 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
5401 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
5461 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
5521 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
5581 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
5641 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
5701 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
5761 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
5821 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
5881 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
5941 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
6001 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
6061 gaaattaacc ctcactaaag ggaacaaaag ctggtaccaa ttccttgaat tttcaaaaat
6121 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
6181 accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
6241 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
6301 gctatattga agta
//
