| 质粒名称: | pQZ210 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 106414 |
| 存储实验室: | Axel Brunger |
|
载体骨架基本信息 |
|
| 载体名称: | pACYCDuet-1 |
| 载体出品公司: | Novagen |
| 载体抗性: | 氯霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 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: | Rat Synaptobrevin-2 fragment 28-66,VAMP2 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | 片段28-66，密码子优化 |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: | His tag（插入N端子） |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | ACYCDuetUP1 Primer #71178-3 |
| 3' 测序引物 | 次 |
|
文献引用方法 |
|
| 材料和方法部分: | pQZ210 was a gift from Axel Brunger (Addgene plasmid # 106414 ; http://n2t.net/addgene:106414 ; RRID:Addgene_106414) |
| 参考文献部分: | The primed SNARE-complexin-synaptotagmin complex for neuronal exocytosis. Zhou Q, Zhou P, Wang AL, Wu D, Zhao M, Sudhof TC, Brunger AT. Nature. 2017 Aug 24;548(7668):420-425. doi: 10.1038/nature23484. Epub 2017 Aug 16. 10.1038/nature23484 PubMed 28813412 |
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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 pQZ210 4349 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Co-expresses trans SNARE complex with pDC183 in E.coli.
ACCESSION .
VERSION .
KEYWORDS pQZ210
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4349)
AUTHORS Zhou Q, Zhou P, Wang AL, Wu D, Zhao M, Sudhof TC, Brunger AT
TITLE The primed SNARE-complexin-synaptotagmin complex for neuronal
exocytosis.
JOURNAL Nature. 2017 Aug 24;548(7668):420-425. doi: 10.1038/nature23484.
Epub 2017 Aug 16.
PUBMED 28813412
REFERENCE 2 (bases 1 to 4349)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4349
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..20
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 21..45
/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)."
CDS 104..133
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 134..154
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
promoter 421..439
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 440..464
/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)."
CDS 723..767
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(805..823)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 819..866
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1087..1746)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1661..1680
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(1747..1849)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(2241..2258)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2375..2920)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(2455..2474)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3145..4227)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 4189..4208
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4228..4305)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt
61 ttgtttaact ttaataagga gatataccat ggcatcgtac taccatcacc atcaccatca
121 ccatcaccat cacgaaaacc tgtattttca gggcagcaac aggagactgc agcagaccca
181 ggcccaggtg gatgaggtgg tggacatcat gagggtgaat gtggacaagg tcctggagcg
241 agaccagaag ctatcggaac tggatgatcg ctaggatgcc ctccaggcag gggcctccca
301 gtttgaaaca agtgcagcca agctcaagcg caaatactgg tagctcgaca agcttgcggc
361 cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat
421 taatacgact cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta
481 gttaagtata agaaggagat atacatatgg ccctcagtga gatcgagacc aggcacagtg
541 agatcatcaa gttggagaac agcatccggg agctacacga tatgttcatg gacatggcca
601 tgctggtgga gagccagggg gagatgattg acaggatcga gtacaatgtg gaacacgctg
661 tggactacgt ggagagggcc gtgtctgaca ccaagaaggc cgtcaagtaa ctcgagtctg
721 gtaaagaaac cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag
781 cttaattaac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
841 ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
901 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
961 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
1021 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
1081 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
1141 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
1201 cttgtcgcct tgcgtataat atttgcccat agtgaaaacg ggggcgaaga agttgtccat
1261 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
1321 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
1381 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
1441 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
1501 cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag
1561 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
1621 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
1681 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
1741 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
1801 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
1861 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
1921 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
1981 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
2041 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
2101 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
2161 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
2221 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
2281 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
2341 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
2401 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
2461 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
2521 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
2581 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
2641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
2701 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
2761 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
2821 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
2881 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
2941 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
3001 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
3061 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
3121 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
3181 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
3241 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
3301 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
3361 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
3421 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
3481 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
3541 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
3601 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
3661 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
3721 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
3781 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
3841 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
3901 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
3961 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
4021 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
4081 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
4141 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
4201 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
4261 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
4321 ctcccttatg cgactcctgc attaggaaa
//
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