pGL4.50载体简介
The pGL4.50[luc2/CMV/Hygro] Vector encodes the luciferase reporter gene luc2 (Photinus pyralis), which has been codon optimized for mammalian expression. This vector also is engineered with fewer consensus regulatory sequences for reduced backgrounds and a decreased risk of anomalous transcription. This vector contains the following features: luc2 reporter gene for expression in mammalian cells; CMV promoter for high translational expression; SV40 late poly(A) signal sequence is positioned downstream of luc2 to provide efficient transcription termination and mRNA polyadenylation; Binding region for RV primer 3 and RV primer 4; Synthetic poly(A) signal/transcription start site; Synthetic Hygromycin B-resistance gene for mammalian cell selection of the plasmid; Plasmid replication origin; Ampr gene for bacterial selection for vector amplification.
LOCUS EU921840 6600 bp DNA linear SYN 28-AUG-2008
DEFINITION Luciferase reporter vector pGL4.50[luc2/CMV/Hygro], complete
sequence.
ACCESSION EU921840
VERSION EU921840.1 GI:197215832
KEYWORDS .
SOURCE Luciferase reporter vector pGL4.50[luc2/CMV/Hygro]
ORGANISM Luciferase reporter vector pGL4.50[luc2/CMV/Hygro]
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 6600)
AUTHORS Brouette,C., Hung,L. and Kopish,K.
TITLE pGL4 Luciferase Reporter Vectors Technical Manual, TM259
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6600)
AUTHORS Brouette,C., Hung,L. and Kopish,K.
TITLE Direct Submission
JOURNAL Submitted (24-JUL-2008) SciComm, Promega, 5500 E Cheryl Parkway,
Madison, WI 53711, USA
FEATURES Location/Qualifiers
source 1..6600
/organism="Luciferase reporter vector
pGL4.50[luc2/CMV/Hygro]"
/mol_type="other DNA"
/db_xref="taxon:554672"
regulatory 14..755
/regulatory_class="promoter"
/note="CMV immediate early enhancer/promoter"
gene 859..2511
/gene="luc2"
CDS 859..2511
/gene="luc2"
/note="luminescent reporter protein"
/codon_start=1
/transl_table=11
/product="firefly luciferase"
/protein_id="ACH53163.1"
/db_xref="GI:197215833"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
AHIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAV
APANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQG
FQSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVR
FSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRS
LQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLP
GIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCV
RGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVA
PAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTT
AKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
misc_feature 2546..2767
/note="SV40 late poly(A) region"
regulatory 2815..3233
/regulatory_class="promoter"
/note="SV40 early enhancer/promoter"
gene 3258..4295
/gene="HygroR"
CDS 3258..4295
/gene="HygroR"
/codon_start=1
/transl_table=11
/product="hygromycin resistant protein"
/protein_id="ACH53164.1"
/db_xref="GI:197215834"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQD
LPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWS
EAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQ
SLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPS
TRPRAKEVGRV"
misc_feature 4319..4367
/note="poly(A) region"
primer_bind complement(4434..4453)
/note="RV primer 4 binding region"
gene complement(5482..6342)
/gene="AmpR"
CDS complement(5482..6342)
/gene="AmpR"
/note="confers resistance to ampicillin"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="ACH53165.1"
/db_xref="GI:197215835"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
misc_feature 6447..6600
/note="poly(A) signal/transcription pause region"
primer_bind 6549..6568
/note="RV primer 3 binding region"
ORIGIN
1 ggcctaactg gcctcaatat tggccattag ccatattatt cattggttat atagcataaa
61 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
121 ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt tattaatagt
181 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
241 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
301 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
361 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
421 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
481 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
541 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
601 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
661 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
721 atataagcag agctcgttta gtgaaccgtc agatcactag aagctttatt gcggtagttt
781 atcacagtta aattgctaac gcagtcagtg ggcctcggcg gccaagcttg gcaatccggt
841 actgttggta aagccaccat ggaagatgcc aaaaacatta agaagggccc agcgccattc
901 tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc
961 ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc
1021 gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca
1081 aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt
1141 gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg
1201 ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa
1261 aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc
1321 aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc
1381 ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg
1441 atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc
1501 gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac
1561 accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc
1621 tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg
1681 cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc
1741 ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc
1801 ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca
1861 ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa
1921 ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg
1981 gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc
2041 cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag
2101 gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc
2161 gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg
2221 gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac
2281 gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc
2341 gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt
2401 ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag
2461 atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta ataattctag
2521 agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa
2581 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
2641 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
2701 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
2761 tgtggtaaaa tcgataagga tccgtttgcg tattgggcgc tcttccgctg atctgcgcag
2821 caccatggcc tgaaataacc tctgaaagag gaacttggtt agctaccttc tgaggcggaa
2881 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
3001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
3061 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
3121 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
3181 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcgattctt
3241 ctgacactag cgccaccatg aagaagcccg aactcaccgc taccagcgtt gaaaaatttc
3301 tcatcgagaa gttcgacagt gtgagcgacc tgatgcagtt gtcggagggc gaagagagcc
3361 gagccttcag cttcgatgtc ggcggacgcg gctatgtact gcgggtgaat agctgcgctg
3421 atggcttcta caaagaccgc tacgtgtacc gccacttcgc cagcgctgca ctacccatcc
3481 ccgaagtgtt ggacatcggc gagttcagcg agagcctgac atactgcatc agtagacgcg
3541 cccaaggcgt tactctccaa gacctccccg aaacagagct gcctgctgtg ttacagcctg
3601 tcgccgaagc tatggatgct attgccgccg ccgacctcag tcaaaccagc ggcttcggcc
3661 cattcgggcc ccaaggcatc ggccagtaca caacctggcg ggatttcatt tgcgccattg
3721 ctgatcccca tgtctaccac tggcagaccg tgatggacga caccgtgtcc gccagcgtag
3781 ctcaagccct ggacgaactg atgctgtggg ccgaagactg tcccgaggtg cgccacctcg
3841 tccatgccga cttcggcagc aacaacgtcc tgaccgacaa cggccgcatc accgccgtaa
3901 tcgactggtc cgaagctatg ttcggggaca gtcagtacga ggtggccaac atcttcttct
3961 ggcggccctg gctggcttgc atggagcagc agactcgcta cttcgagcgc cggcatcccg
4021 agctggccgg cagccctcgt ctgcgagcct acatgctgcg catcggcctg gatcagctct
4081 accagagcct cgtggacggc aacttcgacg atgctgcctg ggctcaaggc cgctgcgatg
4141 ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca aatcgctcgc cggagcgcag
4201 ccgtatggac cgacggctgc gtcgaggtgc tggccgacag cggcaaccgc cggcccagta
4261 cacgaccgcg cgctaaggag gtaggtcgag tttaaactct agaaccggtc atggccgcaa
4321 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgttc gaactagatg
4381 ctgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
4441 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
4501 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
4561 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
4621 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
4681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
4741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
4801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
4861 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
4921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
4981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
5041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
5101 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
5161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
5221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
5281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
5341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
5401 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agcggccgca
5461 aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga tctcagcgat
5521 ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca ctacgattcg
5581 tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc gttcaccggc
5641 ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa gtggtcctgc
5701 tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag taagaagttc
5761 gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg tatcacgctc
5821 gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg tcacatgatc
5881 acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg tcagaagtaa
5941 gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc ttaccgtcat
6001 gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt tttgtgagta
6061 gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca ccgcgccaca
6121 tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga aagactcaag
6181 gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca gttgatcttc
6241 agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc
6301 aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc tttttcaata
6361 ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa gtaatattaa
6421 ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta catctgtgtg
6481 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
6541 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//