| 质粒名称: | pAraGFPCDF |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 47516 |
| 存储实验室: | Bryan Berger |
|
载体骨架基本信息 |
|
| 载体名称: | pCDF |
| 完整质粒大小: | 4939 bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 奇霉素 |
| 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: | GFP |
|
文献引用方法 |
|
| 材料和方法部分: | pAraGFPCDF was a gift from Bryan Berger (Addgene plasmid # 47516 ; http://n2t.net/addgene:47516 ; RRID:Addgene_47516) |
| 参考文献部分: | A Novel Assay for Assessing Juxtamembrane and Transmembrane Domain Interactions Important for Receptor Heterodimerization. Su PC, Berger BW. J Mol Biol. 2013 Jul 19. pii: S0022-2836(13)00456-7. doi: 10.1016/j.jmb.2013.07.022. 10.1016/j.jmb.2013.07.022 PubMed 23876708 |
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载体基本信息
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载体简介
载体序列
LOCUS pAraGFPCDF 4939 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Contains PBAD::GFP reporter, used for heterodimer competition assay.
ACCESSION .
VERSION .
KEYWORDS pAraGFPCDF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4939)
AUTHORS Su PC, Berger BW
TITLE A Novel Assay for Assessing Juxtamembrane and Transmembrane Domain
Interactions Important for Receptor Heterodimerization.
JOURNAL J Mol Biol. 2013 Jul 19. pii: S0022-2836(13)00456-7. doi:
10.1016/j.jmb.2013.07.022.
PUBMED 23876708
REFERENCE 2 (bases 1 to 4939)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4939
/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)."
CDS 83..100
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter 221..505
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 428..447
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 566..1282
/codon_start=1
/product="Aequoria victoria green fluorescent protein"
/label=GFP
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind 1219..1239
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
promoter 1372..1390
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1391..1415
/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 1524..1568
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(1606..1624)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1620..1667
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1838..2629)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(2630..2721)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2769..3507)
/direction=LEFT
/label=CloDF13 ori
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
CDS complement(3717..4799)
/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 4761..4780
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4800..4877)
/gene="lacI"
/label=lacI promoter
promoter 4923..2
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcatc
121 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
181 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
241 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
301 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
361 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
421 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
481 tcgcaactct ctactgtttc tccatacccg tttttttggg aaaaaaaccc ggggatcctc
541 tagatttaat gaaggagata tacatatgag taaaggagaa gaacttttca ctggagtggt
601 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
661 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
721 actacctgtt ccatggccaa cacttgtcac tactttcgcg tatggtcttc aatgctttgc
781 gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta
841 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
901 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
961 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
1021 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattaa
1081 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
1141 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
1201 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
1261 catggatgaa ctatacaaat aattttttct gcaggtcgac aagcttgcgg ccgcataatg
1321 cttaagtcga acagaaagta atcgtattgt acacggccgc ataatcgaaa ttaatacgac
1381 tcactatagg ggaattgtga gcggataaca attccccatc ttagtatatt agttaagtat
1441 aagaaggaga tatacatatg gcagatctca attggatatc ggccggccac gcgatcgctg
1501 acgtcggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
1561 tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac
1621 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
1681 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
1741 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtca tcgtggccgg
1801 atcttgcggc ccctcggctt gaacgaattg ttagacatta tttgccgact accttggtga
1861 tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat
1921 cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg
1981 gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg
2041 cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg
2101 gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa
2161 ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt
2221 ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa
2281 tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa
2341 tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag
2401 gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc
2461 ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg
2521 cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa
2581 ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata ctcttccttt
2641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
2701 gtatttagaa aaataaacaa atagctagct cactcggtcg ctacgctccg ggcgtgagac
2761 tgcggcgggc gctgcggaca catacaaagt tacccacaga ttccgtggat aagcagggga
2821 ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg tttttccata ggctccgccc
2881 tcctgccaga gttcacataa acagacgctt ttccggtgca tctgtgggag ccgtgaggct
2941 caaccatgaa tctgacagta cgggcgaaac ccgacaggac ttaaagatcc ccaccgtttc
3001 cggcgggtcg ctccctcttg cgctctcctg ttccgaccct gccgtttacc ggatacctgt
3061 tccgcctttc tcccttacgg gaagtgtggc gctttctcat agctcacaca ctggtatctc
3121 ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc aagaactccc cgttcagccc
3181 gactgctgcg ccttatccgg taactgttca cttgagtcca acccggaaaa gcacggtaaa
3241 acgccactgg cagcagccat tggtaactgg gagttcgcag aggatttgtt tagctaaaca
3301 cgcggttgct cttgaagtgt gcgccaaagt ccggctacac tggaaggaca gatttggttg
3361 ctgtgctctg cgaaagccag ttaccacggt taagcagttc cccaactgac ttaaccttcg
3421 atcaaaccac ctccccaggt ggttttttcg tttacagggc aaaagattac gcgcagaaaa
3481 aaaggatctc aagaagatcc tttgatcttt tctactgaac cgctctagat ttcagtgcaa
3541 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
3601 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
3661 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
3721 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
3781 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
3841 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
3901 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
3961 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
4021 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
4081 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
4141 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
4201 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
4261 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
4321 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
4381 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
4441 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
4501 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
4561 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
4621 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
4681 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
4741 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
4801 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
4861 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
4921 attaatacga ctcactata
//
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