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pGL4.78[hRlucCP/Hygro]
大量
钰博生物
1ug/5ug
出品公司: | Ybscience |
---|---|
载体名称: | pGL4.78 ; pGL4.78[hRlucCP/Hygro] |
质粒类型: | 信号通路报告载体;哺乳动物载体;海肾荧光素酶报告载体 |
高拷贝/低拷贝: | -- |
克隆方法: | 限制性内切酶,多克隆位点 |
启动子: | 无 |
载体大小: | 5303 bp |
5' 测序引物及序列: | -- |
3' 测序引物及序列: | -- |
载体标签: | hRluc |
载体抗性: | 氨苄青霉素 |
筛选标记: | 潮霉素(Hygromycin) |
克隆菌株: | TOP10等常规菌株 |
宿主细胞(系): | HEK293等 |
备注: | pGL4.78[hRlucCP/Hygro]载体是信号通路报告载体;无启动子;含海肾荧光素酶报告基因hRlucCP。 |
稳定性: | 稳表达 |
组成型/诱导型: | 组成型 |
病毒/非病毒: | 非病毒 |
The pGL4.78[hRlucCP/Hygro] Vector encodes the luciferase reporter gene hRlucCP (Renilla reniformis) and is designed for high expression and reduced anomalous transcription. This vector also contains a mammalian selectable marker for hygromycin resistance in which the number of transcription factor binding sites has been reduced and mammalian codon usage optimized. The pGL4 Vectors are engineered with fewer consensus regulatory sequences than the pGL3 Vectors and a synthetic reporter gene that has been codon optimized for mammalian expression. The pGL4.78[hRlucCP/Hygro] Vector is a basic vector with no promoter. However, it contains a multiple cloning region that allows cloning of a promoter of choice. The hRlucCP reporter gene contains two protein destabilization sequences, hCL1 and hPEST. The protein encoded by hRlucCP responds more quickly and with greater magnitude to changes in transcriptional activity than the hRluc gene, its more stable counterpart.
LOCUS AY864933 5303 bp DNA circular SYN 31-JAN-2005DEFINITION Luciferase reporter vector pGL4.78[hRlucCP/Hygro], complete sequence.ACCESSION AY864933VERSION AY864933.1 GI:58201885KEYWORDS .SOURCE Luciferase reporter vector pGL4.78[hRlucCP/Hygro] ORGANISM Luciferase reporter vector pGL4.78[hRlucCP/Hygro] other sequences; artificial sequences; vectors.REFERENCE 1 (bases 1 to 5303) AUTHORS Paguio,A., Almond,B. and Browning,J. TITLE pGL4 Luciferase Reporter Vectors Technical Manual, TM259 JOURNAL UnpublishedREFERENCE 2 (bases 1 to 5303) AUTHORS Paguio,A., Almond,B. and Browning,J. TITLE Direct Submission JOURNAL Submitted (28-DEC-2004) Scientific Communications, Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USAFEATURES Location/Qualifiers source 1..5303 /organism="Luciferase reporter vector pGL4.78[hRlucCP/Hygro]" /mol_type="genomic DNA" /db_xref="taxon:309059" misc_feature 1..70 /note="multiple cloning site" CDS 100..1212 /note="Renilla luciferase" /codon_start=1 /transl_table=11 /product="luciferase hRlucCP" /protein_id="AAW66997.1" /db_xref="GI:58201886" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAE NAVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLT AWFELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIE EDIALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLS WPREIPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNT EFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQNSACKNWFSSLSHFVIHLNSHGF PPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV" misc_feature 1249..1470 /note="SV40 late poly A" misc_feature 1518..1936 /note="SV40 enhancer/promoter" CDS 1961..2998 /codon_start=1 /transl_table=11 /product="synthetic hygromycin resistance protein" /protein_id="AAW66998.1" /db_xref="GI:58201887" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS TRPRAKEVGRV" regulatory 3022..3070 /regulatory_class="polyA_signal_sequence" /note="synthetic polyadenylation signal sequence" primer_bind 3137..3156 /note="RVprimer4 binding site" rep_origin 3394 /note="ColEI-derived plasmid replication origin" CDS complement(4185..5045) /note="Ampr" /codon_start=1 /transl_table=11 /product="synthetic beta-lactamase" /protein_id="AAW66999.1" /db_xref="GI:58201888" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" misc_feature 5150..5303 /note="synthetic poly(A) signal/transcriptional pause site" primer_bind 5252..5271 /note="RVprimer3 binding site"ORIGIN 1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaagatc tggcctcggc 61 ggccaagctt ggcaatccgg tactgttggt aaagccacca tggcttccaa ggtgtacgac 121 cccgagcaac gcaaacgcat gatcactggg cctcagtggt gggctcgctg caagcaaatg 181 aacgtgctgg actccttcat caactactat gattccgaga agcacgccga gaacgccgtg 241 atttttctgc atggtaacgc tgcctccagc tacctgtgga ggcacgtcgt gcctcacatc 301 gagcccgtgg ctagatgcat catccctgat ctgatcggaa tgggtaagtc cggcaagagc 361 gggaatggct catatcgcct cctggatcac tacaagtacc tcaccgcttg gttcgagctg 421 ctgaaccttc caaagaaaat catctttgtg ggccacgact ggggggcttg tctggccttt 481 cactactcct acgagcacca agacaagatc aaggccatcg tccatgctga gagtgtcgtg 541 gacgtgatcg agtcctggga cgagtggcct gacatcgagg aggatatcgc cctgatcaag 601 agcgaagagg gcgagaaaat ggtgcttgag aataacttct tcgtcgagac catgctccca 661 agcaagatca tgcggaaact ggagcctgag gagttcgctg cctacctgga gccattcaag 721 gagaagggcg aggttagacg gcctaccctc tcctggcctc gcgagatccc tctcgttaag 781 ggaggcaagc ccgacgtcgt ccagattgtc cgcaactaca acgcctacct tcgggccagc 841 gacgatctgc ctaagatgtt catcgagtcc gaccctgggt tcttttccaa cgctattgtc 901 gagggagcta agaagttccc taacaccgag ttcgtgaagg tgaagggcct ccacttcagc 961 caggaggacg ctccagatga aatgggtaag tacatcaaga gcttcgtgga gcgcgtgctg 1021 aagaacgagc agaattctgc ttgcaagaac tggttcagta gcttaagcca ctttgtgatc 1081 caccttaaca gccacggctt ccctcccgag gtggaggagc aggccgccgg caccctgccc 1141 atgagctgcg cccaggagag cggcatggat agacaccctg ctgcttgcgc cagcgccagg 1201 atcaacgtct aactgcagtc tagagtcggg gcggccggcc gcttcgagca gacatgataa 1261 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt 1321 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta 1381 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt 1441 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgttt gcgtattggg 1501 cgctcttccg ctgatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg 1561 gttagctacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg 1621 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 1681 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 1741 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 1801 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 1861 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 1921 cttttgcaaa aagctcgatt cttctgacac tagcgccacc atgaagaagc ccgaactcac 1981 cgctaccagc gttgaaaaat ttctcatcga gaagttcgac agtgtgagcg acctgatgca 2041 gttgtcggag ggcgaagaga gccgagcctt cagcttcgat gtcggcggac gcggctatgt 2101 actgcgggtg aatagctgcg ctgatggctt ctacaaagac cgctacgtgt accgccactt 2161 cgccagcgct gcactaccca tccccgaagt gttggacatc ggcgagttca gcgagagcct 2221 gacatactgc atcagtagac gcgcccaagg cgttactctc caagacctcc ccgaaacaga 2281 gctgcctgct gtgttacagc ctgtcgccga agctatggat gctattgccg ccgccgacct 2341 cagtcaaacc agcggcttcg gcccattcgg gccccaaggc atcggccagt acacaacctg 2401 gcgggatttc atttgcgcca ttgctgatcc ccatgtctac cactggcaga ccgtgatgga 2461 cgacaccgtg tccgccagcg tagctcaagc cctggacgaa ctgatgctgt gggccgaaga 2521 ctgtcccgag gtgcgccacc tcgtccatgc cgacttcggc agcaacaacg tcctgaccga 2581 caacggccgc atcaccgccg taatcgactg gtccgaagct atgttcgggg acagtcagta 2641 cgaggtggcc aacatcttct tctggcggcc ctggctggct tgcatggagc agcagactcg 2701 ctacttcgag cgccggcatc ccgagctggc cggcagccct cgtctgcgag cctacatgct 2761 gcgcatcggc ctggatcagc tctaccagag cctcgtggac ggcaacttcg acgatgctgc 2821 ctgggctcaa ggccgctgcg atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac 2881 acaaatcgct cgccggagcg cagccgtatg gaccgacggc tgcgtcgagg tgctggccga 2941 cagcggcaac cgccggccca gtacacgacc gcgcgctaag gaggtaggtc gagtttaaac 3001 tctagaaccg gtcatggccg caataaaata tctttatttt cattacatct gtgtgttggt 3061 tttttgtgtg ttcgaactag atgctgtcga ccgatgccct tgagagcctt caacccagtc 3121 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt 3181 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc 3241 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 3301 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 3361 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 3421 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 3481 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 3541 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 3601 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 3661 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 3721 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 3781 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 3841 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 3901 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 3961 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 4021 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 4081 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 4141 aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca gtgcttgatc 4201 agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc ctgactcccc 4261 gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc agcaatgatg 4321 ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg 4381 gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat gagctgctgt 4441 cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt ggccattgct 4501 actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc tggttcccag 4561 cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag ctccttaggg 4621 cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt aatggcagca 4681 ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac cggcgagtac 4741 tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg cccggcgtct 4801 atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat cgggaatcgt 4861 tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc gatatagccc 4921 actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc ggggtgtgca 4981 aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg 5041 ctcatactcg tcctttttca atattattga agcatttatc agggttacta gtacgtctct 5101 caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca ataaaatatc 5161 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc 5221 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag 5281 tgcaggtgcc agaacatttc tct
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