| 质粒名称: | ELONGIN BC (XX01TCEB1A-c001) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 110274 |
| 存储实验室: | Nicola Burgess-Brown |
|
载体骨架基本信息 |
|
| 载体名称: | pACYC-Duet1 |
| 载体出品公司: | Novagen |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氯霉素25 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Elongin BC,co-expresses Elongin B and Elongin C |
| 物种来源: | H. sapiens (human) |
| 基因突变: | Elongin B is full length. Elongin C contains a.a. 17-112. |
| 启动子: | T7 |
| 5’测序引物 | pDuetDOWN-1 (5'-TTGTACACGGCCGCATAATC-3') or pDuetUP2 (5'-GATTATGCGGCCGTGTACAA-3') |
| 3’测序引物 | T7-term |
|
文献引用方法 |
|
| 材料和方法部分: | ELONGIN BC (XX01TCEB1A-c001) was a gift from Nicola Burgess-Brown (Addgene plasmid # 110274 ; http://n2t.net/addgene:110274 ; RRID:Addgene_110274) |
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 ELONGIN BC (XX01TCEB1A-c001) 4587 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Elongin B and Elongin C co-expression plasmid for structure
determination; Full length Elongin B.
ACCESSION .
VERSION .
KEYWORDS ELONGIN BC (XX01TCEB1A-c001)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4587)
TITLE SGC Oxford April 2018 plasmids
REFERENCE 2 (bases 1 to 4587)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4587
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3..27
/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)."
promoter 502..520
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 521..545
/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 943..987
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(1025..1043)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1039..1086
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1307..1966)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1881..1900
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(1967..2069)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(2461..2478)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2595..3140)
/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(2675..2694)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3365..4447)
/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 4409..4428
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4448..4525)
/gene="lacI"
/label=lacI promoter
promoter 4571..2
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atggacgtgt ttctcatgat ccggcgccac aagaccacca tcttcacgga
121 cgccaaggag tccagcacgg tgttcgaact gaagcgcatc gtcgagggca tcctcaagcg
181 gcctcctgac gagcagcggc tgtacaagga tgaccaactc ttggatgatg gcaagacact
241 gggcgagtgt ggcttcacca gtcaaacagc acggccacag gccccagcca cagtggggct
301 ggccttccgg gcagatgaca ccttcgaggc cctgtgcatc gagccgtttt ccagcccgcc
361 agagctgccc gatgtgatga agccccagga ctcgggaagc agtgccaatg aacaagctgt
421 gcagtagggg aagcttgcgg ccgcataatg cttaagtcga acagaaagta atcgtattgt
481 acacggccgc ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
541 attccccatc ttagtatatt agttaagtat aagaaggaga tatacatatg atgtatgtca
601 aattgatatc atctgatggc catgaattta ttgtaaaaag agaacatgca ttaacatcag
661 gcacgataaa agccatgttg agtggcccag gtcagtttgc tgagaacgaa accaatgagg
721 tcaattttag agagatacct tcacatgtgc tatcgaaagt atgcatgtat tttacgtaca
781 aggttcgcta cactaacagc tccaccgaga ttcctgaatt cccaattgca cctgaaattg
841 cactggaact gctgatggct gcgaacttct tagattgtta gggatctcaa ttggatatcg
901 gccggccacg cgatcgctga cgtcggtacc ctcgagtctg gtaaagaaac cgctgctgcg
961 aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac ctaggctgct
1021 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
1081 tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
1141 aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg
1201 accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
1261 cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
1321 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
1381 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
1441 atttgcccat agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
1501 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
1561 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
1621 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
1681 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
1741 ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
1801 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
1861 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
1921 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
1981 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
2041 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
2101 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
2161 caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta
2221 tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca
2281 gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt
2341 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
2401 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
2461 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
2521 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
2581 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
2641 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc
2701 ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta
2761 tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa
2821 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta
2881 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa
2941 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa
3001 gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga
3061 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc
3121 gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt
3181 cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt
3241 aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
3301 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
3361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
3421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
3481 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
3541 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
3601 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc
3661 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
3721 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
3781 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
3841 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
3901 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
3961 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
4021 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
4081 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
4141 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
4201 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
4261 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
4321 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
4381 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
4441 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
4501 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
4561 attaggaaat taatacgact cactata
//
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