pGL4.76载体简介
The pGL4.76[hRluc/Hygro] Vector encodes the luciferase reporter gene hRluc (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 pGL4.76[hRluc/Hygro] Vector is a basic vector with no promoter. However, it contains a multiple cloning region that allows cloning of a promoter of choice.
LOCUS AY864931 5121 bp DNA circular SYN 31-JAN-2005
DEFINITION Luciferase reporter vector pGL4.76[hRluc/Hygro], complete sequence.
ACCESSION AY864931
VERSION AY864931.1 GI:58201877
KEYWORDS .
SOURCE Luciferase reporter vector pGL4.76[hRluc/Hygro]
ORGANISM Luciferase reporter vector pGL4.76[hRluc/Hygro]
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 5121)
AUTHORS Paguio,A., Almond,B. and Browning,J.
TITLE pGL4 Luciferase Reporter Vectors Technical Manual, TM259
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5121)
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, USA
FEATURES Location/Qualifiers
source 1..5121
/organism="Luciferase reporter vector
pGL4.76[hRluc/Hygro]"
/mol_type="genomic DNA"
/db_xref="taxon:309057"
misc_feature 1..70
/note="multiple cloning site"
CDS 100..1035
/note="Renilla luciferase"
/codon_start=1
/transl_table=11
/product="luciferase hRluc"
/protein_id="AAW66991.1"
/db_xref="GI:58201878"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAE
NAVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLT
AWFELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIE
EDIALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLS
WPREIPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNT
EFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
misc_feature 1067..1288
/note="SV40 late poly A"
misc_feature 1336..1754
/note="SV40 enhancer/promoter"
CDS 1779..2816
/codon_start=1
/transl_table=11
/product="synthetic hygromycin resistance protein"
/protein_id="AAW66992.1"
/db_xref="GI:58201879"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
TRPRAKEVGRV"
regulatory 2840..2888
/regulatory_class="polyA_signal_sequence"
/note="synthetic polyadenylation signal sequence"
primer_bind 2955..2974
/note="RVprimer4 binding site"
rep_origin 3212
/note="ColEI-derived plasmid replication origin"
CDS complement(4003..4863)
/note="Ampr"
/codon_start=1
/transl_table=11
/product="synthetic beta-lactamase"
/protein_id="AAW66993.1"
/db_xref="GI:58201880"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
misc_feature 4968..5121
/note="synthetic poly(A) signal/transcriptional pause
site"
primer_bind 5070..5089
/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 agtaattcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
1081 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
1141 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
1201 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
1261 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtttgc gtattgggcg
1321 ctcttccgct gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
1381 tagctacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
1441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
1501 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
1561 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
1621 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
1681 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
1741 tttgcaaaaa gctcgattct tctgacacta gcgccaccat gaagaagccc gaactcaccg
1801 ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac ctgatgcagt
1861 tgtcggaggg cgaagagagc cgagccttca gcttcgatgt cggcggacgc ggctatgtac
1921 tgcgggtgaa tagctgcgct gatggcttct acaaagaccg ctacgtgtac cgccacttcg
1981 ccagcgctgc actacccatc cccgaagtgt tggacatcgg cgagttcagc gagagcctga
2041 catactgcat cagtagacgc gcccaaggcg ttactctcca agacctcccc gaaacagagc
2101 tgcctgctgt gttacagcct gtcgccgaag ctatggatgc tattgccgcc gccgacctca
2161 gtcaaaccag cggcttcggc ccattcgggc cccaaggcat cggccagtac acaacctggc
2221 gggatttcat ttgcgccatt gctgatcccc atgtctacca ctggcagacc gtgatggacg
2281 acaccgtgtc cgccagcgta gctcaagccc tggacgaact gatgctgtgg gccgaagact
2341 gtcccgaggt gcgccacctc gtccatgccg acttcggcag caacaacgtc ctgaccgaca
2401 acggccgcat caccgccgta atcgactggt ccgaagctat gttcggggac agtcagtacg
2461 aggtggccaa catcttcttc tggcggccct ggctggcttg catggagcag cagactcgct
2521 acttcgagcg ccggcatccc gagctggccg gcagccctcg tctgcgagcc tacatgctgc
2581 gcatcggcct ggatcagctc taccagagcc tcgtggacgg caacttcgac gatgctgcct
2641 gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc ggtcgcacac
2701 aaatcgctcg ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg ctggccgaca
2761 gcggcaaccg ccggcccagt acacgaccgc gcgctaagga ggtaggtcga gtttaaactc
2821 tagaaccggt catggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
2881 tttgtgtgtt cgaactagat gctgtcgacc gatgcccttg agagccttca acccagtcag
2941 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
3001 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
3061 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
3121 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
3181 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
3241 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
3301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
3361 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
3421 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
3481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
3541 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
3601 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
3661 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
3721 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
3781 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
3841 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
3901 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
3961 cttggtctga cagcggccgc aaatgctaaa ccactgcagt ggttaccagt gcttgatcag
4021 tgaggcaccg atctcagcga tctgcctatt tcgttcgtcc atagtggcct gactccccgt
4081 cgtgtagatc actacgattc gtgagggctt accatcaggc cccagcgcag caatgatgcc
4141 gcgagagccg cgttcaccgg cccccgattt gtcagcaatg aaccagccag cagggagggc
4201 cgagcgaaga agtggtcctg ctactttgtc cgcctccatc cagtctatga gctgctgtcg
4261 tgatgctaga gtaagaagtt cgccagtgag tagtttccga agagttgtgg ccattgctac
4321 tggcatcgtg gtatcacgct cgtcgttcgg tatggcttcg ttcaactctg gttcccagcg
4381 gtcaagccgg gtcacatgat cacccatatt atgaagaaat gcagtcagct ccttagggcc
4441 tccgatcgtt gtcagaagta agttggccgc ggtgttgtcg ctcatggtaa tggcagcact
4501 acacaattct cttaccgtca tgccatccgt aagatgcttt tccgtgaccg gcgagtactc
4561 aaccaagtcg ttttgtgagt agtgtatacg gcgaccaagc tgctcttgcc cggcgtctat
4621 acgggacaac accgcgccac atagcagtac tttgaaagtg ctcatcatcg ggaatcgttc
4681 ttcggggcgg aaagactcaa ggatcttgcc gctattgaga tccagttcga tatagcccac
4741 tcttgcaccc agttgatctt cagcatcttt tactttcacc agcgtttcgg ggtgtgcaaa
4801 aacaggcaag caaaatgccg caaagaaggg aatgagtgcg acacgaaaat gttggatgct
4861 catactcgtc ctttttcaat attattgaag catttatcag ggttactagt acgtctctca
4921 aggataagta agtaatatta aggtacggga ggtattggac aggccgcaat aaaatatctt
4981 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc
5041 tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg
5101 caggtgccag aacatttctc t
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