pGL4.80载体简介
The pGL4.80[hRlucP/Neo] Vector encodes the luciferase reporter gene hRlucP (Renilla reniformis) and is designed for high expression and reduced anomalous transcription. This vector also contains a mammalian selectable marker for neomycin 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 pGL4.80[hRlucP/Neo] Vector is a basic vector with no promoter. However, it contains a multiple cloning region that allows cloning of a promoter of choice. The hRlucP reporter gene contains hPEST, a protein destabilization sequence. The protein encoded by hRlucP responds more quickly and with a greater magnitude to changes in transcriptional activity than the hRluc gene, its more stable counterpart.
LOCUS DQ188844 5010 bp DNA circular SYN 16-NOV-2005
DEFINITION Synthetic Renilla luciferase reporter vector pGL4.80[hRlucP/Neo],
complete sequence.
ACCESSION DQ188844
VERSION DQ188844.1 GI:76364306
KEYWORDS .
SOURCE Renilla luciferase reporter vector pGL4.80[hRlucP/Neo]
ORGANISM Renilla luciferase reporter vector pGL4.80[hRlucP/Neo]
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 5010)
AUTHORS Stecha,P., Paguio,A. and Chauvin,F.
TITLE pGL4 Luciferase Reporter Vectors Technical Manual, TM259, Promega
Corporation
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5010)
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, USA
FEATURES Location/Qualifiers
source 1..5010
/organism="Renilla luciferase reporter vector
pGL4.80[hRlucP/Neo]"
/mol_type="other DNA"
/db_xref="taxon:346091"
misc_feature 1..70
/note="multiple cloning region"
CDS 100..1158
/note="synthetic Renilla luciferase; contains hPEST
destabilization sequence"
/codon_start=1
/product="hRlucP"
/protein_id="ABA41674.1"
/db_xref="GI:76364307"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAE
NAVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLT
AWFELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIE
EDIALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLS
WPREIPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNT
EFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQNSHGFPPEVEEQAAGTLPMSCAQ
ESGMDRHPAACASARINV"
regulatory 1198..1419
/regulatory_class="polyA_signal_sequence"
/note="SV40 late poly(A) signal"
regulatory 1467..1885
/regulatory_class="promoter"
/note="SV40 early enhancer/promoter"
CDS 1910..2704
/note="neomycin resistance"
/codon_start=1
/product="neomycin phosphotransferase"
/protein_id="ABA41675.1"
/db_xref="GI:76364308"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
regulatory 2729..2777
/regulatory_class="polyA_signal_sequence"
/note="synthetic poly(A) signal"
primer_bind 2844..2863
/note="Reporter Vector primer 4 (RVprimer4) binding
region"
rep_origin 3101
/note="ColEI-derived plasmid replication origin"
gene complement(3892..4752)
/gene="bla"
CDS complement(3892..4752)
/gene="bla"
/note="ampicillin resistance"
/codon_start=1
/product="beta-lactamase"
/protein_id="ABA41676.1"
/db_xref="GI:76364309"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
regulatory 4857..5010
/regulatory_class="polyA_signal_sequence"
/note="synthetic poly(A) signal/transcriptional pause
site"
primer_bind 4959..4978
/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 agaattctca cggcttccct cccgaggtgg aggagcaggc cgccggcacc
1081 ctgcccatga gctgcgccca ggagagcggc atggatagac accctgctgc ttgcgccagc
1141 gccaggatca acgtctaagg ccgcgactct agagtcgggg cggccggccg cttcgagcag
1201 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
1261 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
1321 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
1381 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtttg
1441 cgtattgggc gctcttccgc tgatctgcgc agcaccatgg cctgaaataa cctctgaaag
1501 aggaacttgg ttagctacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
1561 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
1621 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
1681 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
1741 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
1801 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
1861 aggcctaggc ttttgcaaaa agctcgattc ttctgacact agcgccacca tgatcgaaca
1921 agacggcctc catgctggca gtcccgcagc ttgggtcgaa cgcttgttcg ggtacgactg
1981 ggcccagcag accatcggat gtagcgatgc ggccgtgttc cgtctaagcg ctcaaggccg
2041 gcccgtgctg ttcgtgaaga ccgacctgag cggcgccctg aacgagcttc aagacgaggc
2101 tgcccgcctg agctggctgg ccaccaccgg cgtaccctgc gccgctgtgt tggatgttgt
2161 gaccgaagcc ggccgggact ggctgctgct gggcgaggtc cctggccagg atctgctgag
2221 cagccacctt gcccccgctg agaaggtttc tatcatggcc gatgcaatgc ggcgcctgca
2281 caccctggac cccgctacct gccccttcga ccaccaggct aagcatcgga tcgagcgtgc
2341 tcggacccgc atggaggccg gcctggtgga ccaggacgac ctggacgagg agcatcaggg
2401 cctggccccc gctgaactgt tcgcccgact gaaagcccgc atgccggacg gtgaggacct
2461 ggttgtcaca cacggagatg cctgcctccc taacatcatg gtcgagaatg gccgcttctc
2521 cggcttcatc gactgcggtc gcctaggagt tgccgaccgc taccaggaca tcgccctggc
2581 cacccgcgac atcgctgagg agcttggcgg cgagtgggcc gaccgcttct tagtcttgta
2641 cggcatcgca gctcccgaca gccagcgcat cgccttctac cgcttgctcg acgagttctt
2701 ttaatgatct agaaccggtc atggccgcaa taaaatatct ttattttcat tacatctgtg
2761 tgttggtttt ttgtgtgttc gaactagatg ctgtcgaccg atgcccttga gagccttcaa
2821 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
2881 cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca
2941 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
3001 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
3061 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
3121 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
3181 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
3241 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
3301 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
3361 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
3421 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
3481 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
3541 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
3601 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
3661 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
3721 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
3781 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
3841 atgagtaaac ttggtctgac agcggccgca aatgctaaac cactgcagtg gttaccagtg
3901 cttgatcagt gaggcaccga tctcagcgat ctgcctattt cgttcgtcca tagtggcctg
3961 actccccgtc gtgtagatca ctacgattcg tgagggctta ccatcaggcc ccagcgcagc
4021 aatgatgccg cgagagccgc gttcaccggc ccccgatttg tcagcaatga accagccagc
4081 agggagggcc gagcgaagaa gtggtcctgc tactttgtcc gcctccatcc agtctatgag
4141 ctgctgtcgt gatgctagag taagaagttc gccagtgagt agtttccgaa gagttgtggc
4201 cattgctact ggcatcgtgg tatcacgctc gtcgttcggt atggcttcgt tcaactctgg
4261 ttcccagcgg tcaagccggg tcacatgatc acccatatta tgaagaaatg cagtcagctc
4321 cttagggcct ccgatcgttg tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat
4381 ggcagcacta cacaattctc ttaccgtcat gccatccgta agatgctttt ccgtgaccgg
4441 cgagtactca accaagtcgt tttgtgagta gtgtatacgg cgaccaagct gctcttgccc
4501 ggcgtctata cgggacaaca ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg
4561 gaatcgttct tcggggcgga aagactcaag gatcttgccg ctattgagat ccagttcgat
4621 atagcccact cttgcaccca gttgatcttc agcatctttt actttcacca gcgtttcggg
4681 gtgtgcaaaa acaggcaagc aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg
4741 ttggatgctc atactcgtcc tttttcaata ttattgaagc atttatcagg gttactagta
4801 cgtctctcaa ggataagtaa gtaatattaa ggtacgggag gtattggaca ggccgcaata
4861 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa
4921 catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca
4981 gtgcaagtgc aggtgccaga acatttctct
//