| 质粒名称: | pLIC-SM-HA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 111188 |
| 存储实验室: | Silvia Moreno |
|
载体骨架基本信息 |
|
| 载体名称: | pLIC |
| 载体出品公司: | Vern Carruthers |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6000 bp |
| 完整质粒大小: | -- |
| 载体修饰: | Modified by Sheiner et al. (PMID: 22144892) |
| 载体类型: | 弓形虫表达 |
| 筛选标记: | 氯霉素 |
| 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 | Endogenous |
| 标签/融合蛋白: | Spaghetti Monster HA Tag (插入C端子) |
| 克隆方法: | Ligation Independent克隆 |
| 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: | pLIC-SM-HA was a gift from Silvia Moreno (Addgene plasmid # 111188 ; http://n2t.net/addgene:111188 ; RRID:Addgene_111188) |
| 参考文献部分: | Tagging of Weakly Expressed Toxoplasma gondii Calcium-Related Genes with High-Affinity Tags. Hortua Triana MA, Marquez-Nogueras KM, Chang L, Stasic AJ, Li C, Spiegel KA, Sharma A, Li ZH, Moreno SNJ. J Eukaryot Microbiol. 2018 Apr 19. doi: 10.1111/jeu.12626. 10.1111/jeu.12626 PubMed 29672999 |
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 pLIC-SM-HA 5940 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION High affinity HA Tagging of weakly expressed genes (optimized for
Toxoplasma gondii).
ACCESSION .
VERSION .
KEYWORDS pLIC-SM-HA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5940)
AUTHORS Hortua Triana MA, Marquez-Nogueras KM, Chang L, Stasic AJ, Li C,
Spiegel KA, Sharma A, Li ZH, Moreno SNJ
TITLE Tagging of Weakly Expressed Toxoplasma gondii Calcium-Related Genes
with High-Affinity Tags.
JOURNAL J Eukaryot Microbiol. 2018 Apr 19. doi: 10.1111/jeu.12626.
PUBMED 29672999
REFERENCE 2 (bases 1 to 5940)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5940
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 38..59
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 74..104
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 112..128
/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 117..139
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 136..152
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 136..152
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 173..191
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 269..295
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 299..325
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 329..355
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 884..910
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 917..943
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 956..982
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 989..1015
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1226..1252
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1256..1282
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1286..1312
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind 1741..1757
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 1742..1758
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 2273..2935
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MHEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFS
SLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVAN
MDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQVGA"
primer_bind complement(2342..2361)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind complement(3253..3269)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3253..3269)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(3301..3320)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3302..3320)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3327..3344)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3327..3343)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3336..3358)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3485..3940
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3572..3591)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3782..3803
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3966..4070
/gene="bla"
/label=AmpR promoter
CDS 4071..4931
/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(4289..4308)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5102..5690
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5591..5610
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5844..5861
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
61 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
121 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc
181 ctcactaaag ggaacaaaag ctggtaccct gtacttccaa tccaatttaa ttaaaattgg
241 aagtggagga cgggaattcc ctaggatgta cccttatgat gtgcccgatt atgctggcta
301 cccttatgat gttcctgatt acgccggata tccgtatgac gtgccagact acgcgggagg
361 tgtgagcaag ggagaggagt tgtttaccgg cgtggtcccc atactggtgg agttggatgg
421 cgacgttaat ggacataaat tctcggtccg cggcgaggga gagggagacg ccaccaatgg
481 caagctgacg cttaagttca tttgcacgac tggaaaattg cctgtcccct ggccgaccct
541 ggtcaccaca ctgggcggag gagtgcagtg cttctcccgt tacccagacc acatgaagca
601 gcacgatttc ttcaagagcg caatgccgga gggatacgtg caagaacgga ctatttcctt
661 caaggatgac ggaacctata agacccgtgc ggaggtcaaa ttcgagggtg acaccctggt
721 gaaccgaatt gaactcaaag gaatcgattt caaggaggat ggaaatattc tgggtcacaa
781 gctggaatac aacttcaaca gccataatgt gtacattacg gctgataagc agaagaacgg
841 cattaaggcc aatttcaaga tccgccacaa cgttgagggt ggatatccct acgacgtgcc
901 cgattatgcg ggcggatatc cgtatgatgt tccagattat gctggtggtg gaggctatcc
961 ctatgatgtc cccgactacg ccggaggtta cccatacgac gtgcccgact acgctggagg
1021 cgatggcagc gtgcagctgg cagatcatta tcaacagaat acccccatag gtgatggccc
1081 cgttctgctt ccagataatc actacctttc cacccagagc gtgctttcga aagacccgaa
1141 cgaaaagcgt gatcacatgg tcctgctgga gtttgtgacc gcggctggaa tcaccctggg
1201 tatggacgaa ctctacaagg gtggctaccc ctacgatgtg ccggattacg ctggatatcc
1261 gtatgacgta ccggactatg ccggttaccc gtacgatgtc ccggactacg cttaacatat
1321 gtagaaaagt tgtaacgtta gtaaacgtaa cacacgggta taatagccat ttaatggcgc
1381 cgtatatttt tcactccccc tctccgatag gaggaagaaa gacgatccgt ctgcgaatgt
1441 atattacatc gaccggccat agtacgagaa gtggtgccgg tattgttctg tccggggggt
1501 gtaccatcct attttttgta tatgtcgttc ttgtcgggga gcaacagccc atcttccggc
1561 tattgtctcg tatcagttgt gaaacagact gcgttttgcc gaaaatgagt cctatgtccg
1621 atggctattt gtaccgcttc gtaccatttc gcttgtcagc tgtaatcaac cgaattcatt
1681 tggcgagaca tgattccgat taaattcttt actgcgcgtg ctcggatcca ctagttctac
1741 tcgaggtcga cggtatcgat aagcttgatg gcgatgcatg tcccgcgttc gtgaaattct
1801 ctgcatcagc ggagtgatca ggaatcatcg tctcagcggg atgacgttgc ggagcaggcc
1861 ggctcgcggt gggcagtcag atgccgaagg cgtaactcag gacggcttgc gctcatcgca
1921 gaacaggggt ggtgcctgca ttgggtgcgg ttggtgatcc tggttggacc ggtggagatg
1981 cgcgcgcacg aaggggatgt gtcagaaaca ttttgtttgt tctctgtgaa cttttagatg
2041 tgttaaaggc ggcgaatatt agcagagagt cctccttgtt ggattctctc ttgaatttcg
2101 ccctttcctt ctctttgcga gtctcgtaga gaacaagcac tcgttcgccg tccctgacga
2161 cgcaacccgc gcagaagaca tccaccaaac ggtgttacac aatcaccttg tgtgaagttc
2221 ttgcggaaaa ctactcgttg gcattttttc ttgaattccc tttttcgaca aaatgcatga
2281 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
2341 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
2401 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
2461 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
2521 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
2581 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
2641 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
2701 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
2761 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
2821 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
2881 cagaatgctt aatgaattac aacagtactg cgatgagtgg caggtcgggg cgtaattaat
2941 caccgttgtg ctcacttctc aaatcgacaa aggaaacaca cttcgtgcag catgtgcccc
3001 attataaaga aactgagttg ttccgttgtg gcttgcaggt gtcacatcca caaaaaccgg
3061 ccgactctaa ataggagtgt ttcgcagcaa gcagcgaaag tttatgactg ggtccgaatc
3121 tctgaacgga tgtgtggcgg acctggctga tgttgatcgc cgtcgacaca cgcgccacat
3181 gggtcaatac acaagacagc tatcagttgt tttagtcgaa ccggttaaca caattcttgc
3241 ccccccgagg gggatccact agttctagag cggccgccac cgcggtggag ctccaattcg
3301 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
3361 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
3421 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
3481 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
3541 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
3601 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
3661 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
3721 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
3781 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
3841 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
3901 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
3961 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
4021 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
4081 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
4141 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
4201 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
4261 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
4321 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
4381 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
4441 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
4501 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
4561 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
4621 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
4681 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
4741 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
4801 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
4861 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
4921 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
4981 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
5041 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
5101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
5161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
5221 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
5281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
5341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
5401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
5461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
5521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
5581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
5641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
5701 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
5761 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
5821 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
5881 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
//
