pGL4.84载体简介The pGL4.84[hRlucCP/Puro] 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 puromycin resistance in which the number of transcription factor binding sites has been reduced and mammalian codon usage optimized. It has also been engineered with fewer consensus regulatory sequences than the pGL3 Vectors and a synthetic reporter gene that has been codon optimized for mammalian expression. The pGL4.84[hRlucCP/Puro] 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 DQ188848 4866 bp DNA circular SYN 16-NOV-2005DEFINITION Synthetic Renilla luciferase reporter vector pGL4.84[hRlucCP/Puro], complete sequence.ACCESSION DQ188848VERSION DQ188848.1 GI:76364322KEYWORDS .SOURCE Renilla luciferase reporter vector pGL4.84[hRlucCP/Puro] ORGANISM Renilla luciferase reporter vector pGL4.84[hRlucCP/Puro] other sequences; artificial sequences; vectors.REFERENCE 1 (bases 1 to 4866) AUTHORS Stecha,P., Paguio,A. and Chauvin,F. TITLE pGL4 Luciferase Reporter Vectors Technical Manual, TM259, Promega Corporation JOURNAL UnpublishedREFERENCE 2 (bases 1 to 4866) AUTHORS Stecha,P., Paguio,A. and Chauvin,F. TITLE Direct Submission JOURNAL Submitted (31-AUG-2005) Scientific Communications, Promega Corporation, 2800 Woods Hollow Rd., Madison, WI 53711, USAFEATURES Location/Qualifiers source 1..4866 /organism="Renilla luciferase reporter vector pGL4.84[hRlucCP/Puro]" /mol_type="other DNA" /db_xref="taxon:346095" misc_feature 1..70 /note="multiple cloning region" CDS 100..1212 /note="synthetic Renilla luciferase; contains hPEST and hCL1 destabilization sequences" /codon_start=1 /product="hRlucCP" /protein_id="ABA41686.1" /db_xref="GI:76364323" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAE NAVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLT AWFELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIE EDIALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLS WPREIPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNT EFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQNSACKNWFSSLSHFVIHLNSHGF PPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV" regulatory 1249..1470 /regulatory_class="other" /note="SV40 late poly(A) signal" regulatory 1518..1936 /regulatory_class="promoter" /note="SV40 early enhancer/promoter" CDS 1961..2560 /note="puromycin resistance" /codon_start=1 /product="puromycin-N-acetyltransferase" /protein_id="ABA41687.1" /db_xref="GI:76364324" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL ETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" regulatory 2585..2633 /regulatory_class="polyA_signal_sequence" /note="synthetic poly(A) signal" primer_bind 2700..2719 /note="Reporter Vector primer 4 (RVprimer4) binding region" rep_origin 2957 /note="ColEI-derived plasmid replication origin" gene complement(3748..4608) /gene="bla" CDS complement(3748..4608) /gene="bla" /note="ampicillin resistance" /codon_start=1 /product="beta-lactamase" /protein_id="ABA41688.1" /db_xref="GI:76364325" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" regulatory 4713..4866 /regulatory_class="polyA_signal_sequence" /note="synthetic poly(A) signal/transcriptional pause site" primer_bind 4815..4834 /note="Reporter Vector primer 3 (RVprimer3) binding region"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 atgaccgagt acaagcctac 1981 cgtgcgcctg gccactcgcg atgatgtgcc ccgcgccgtc cgcactctgg ccgccgcttt 2041 cgccgactac cccgctaccc ggcacaccgt ggaccccgac cggcacatcg agcgtgtgac 2101 agagttgcag gagctgttcc tgacccgcgt cgggctggac atcggcaagg tgtgggtagc 2161 cgacgacggc gcggccgtgg ccgtgtggac tacccccgag agcgttgagg ccggcgccgt 2221 gttcgccgag atcggccccc gaatggccga gctgagcggc agccgcctgg ccgcccagca 2281 gcaaatggag ggcctgcttg ccccccatcg tcccaaggag cctgcctggt ttctggccac 2341 tgtaggagtg agccccgacc accagggcaa gggcttgggc agcgccgtcg tgttgcccgg 2401 cgtagaggcc gccgaacgcg ccggtgtgcc cgcctttctc gaaacaagcg caccaagaaa 2461 ccttccattc tacgagcgcc tgggcttcac cgtgaccgcc gatgtcgagg tgcccgaggg 2521 acctaggacc tggtgtatga cacgaaaacc tggcgcctaa tgatctagaa ccggtcatgg 2581 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgttcgaac 2641 tagatgctgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc 2701 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg 2761 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 2821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 2881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 2941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 3001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 3061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 3121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 3181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 3241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 3301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 3361 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 3421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 3481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 3541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 3601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 3661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagcg 3721 gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg atcagtgagg caccgatctc 3781 agcgatctgc ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt agatcactac 3841 gattcgtgag ggcttaccat caggccccag cgcagcaatg atgccgcgag agccgcgttc 3901 accggccccc gatttgtcag caatgaacca gccagcaggg agggccgagc gaagaagtgg 3961 tcctgctact ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag 4021 aagttcgcca gtgagtagtt tccgaagagt tgtggccatt gctactggca tcgtggtatc 4081 acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac 4141 atgatcaccc atattatgaa gaaatgcagt cagctcctta gggcctccga tcgttgtcag 4201 aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca gcactacaca attctcttac 4261 cgtcatgcca tccgtaagat gcttttccgt gaccggcgag tactcaacca agtcgttttg 4321 tgagtagtgt atacggcgac caagctgctc ttgcccggcg tctatacggg acaacaccgc 4381 gccacatagc agtactttga aagtgctcat catcgggaat cgttcttcgg ggcggaaaga 4441 ctcaaggatc ttgccgctat tgagatccag ttcgatatag cccactcttg cacccagttg 4501 atcttcagca tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa 4561 tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt 4621 tcaatattat tgaagcattt atcagggtta ctagtacgtc tctcaaggat aagtaagtaa 4681 tattaaggta cgggaggtat tggacaggcc gcaataaaat atctttattt tcattacatc 4741 tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa 4801 cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat 4861 ttctct//