Reporter vector p2luc双Luc荧光素酶报告质粒载体-BioVector NTCC保藏中心
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- 货 号：Reporter vector p2luc
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Reporter vector p2luc质粒载体-BioVector NTCC保藏中心
Order ID | Name | Description |
BiovectorP2LUC | p2Luc reporter vector | p2Luc, 15uL DNA. AmpR.Storage:-20℃ |
Standard heat-shocktransformation of chemically competent bacteria
Take competent cells out of -80°C and thaw on ice.
Take agar plates (containing the appropriate antibiotic) out of 4°C to warm up to room temperature or place in 37°C incubator.
Mix 5μl of DNA into 100μL competent cells in a microcentrifuge or falcon tube. GENTLY mix by flicking the bottom of the tube with your finger a few times.
Place the competent cell/DNA mixture on ice for 20-30min.
Heat shock each transformation tube by placing the bottom 2/3 of the tube into a 42°C water bath for 42 seconds. Put the tubes back on ice for 2 min.
Add 250-500μl SOC media (without antibiotic) and grow in 37°C shaking incubator for 45min.
Plate all of the transformation onto a 250px LB agar plate containing the appropriate antibiotic.
8. Incubate plates at 37°C overnight.
Sequence
LOCUS p2luc 5405 bp ds-DNA circular 18-AUG-2009
DEFINITION Reporter vector p2luc, complete sequence.
ACCESSION AF043450
VERSION AF043450.1 GI:3123919
KEYWORDS .
SOURCE Reporter vector p2luc
ORGANISM Reporter vector p2luc other sequences; artificial sequences;
vectors.
REFERENCE 1 (bases 1 to 5405)
AUTHORS Grentzmann,G., Ingram,J.A., Kelly,P.J., Gesteland,R.F. and
Atkins,J.F.
TITLE A dual-luciferase reporter system for studying recoding signals
JOURNAL RNA 4 (4), 479-486 (1998)
PUBMED 9630253
REFERENCE 2 (bases 1 to 5405)
AUTHORS Grentzmann,G., Gesteland,R.F. and Atkins,J.F.
TITLE Direct Submission
JOURNAL Submitted (16-JAN-1998) HHMI, University of Utah, 6100 Eccles
Bldg., Salt Lake City, UT 84112,USA
COMMENT
COMMENT ApEinfo:methylated:1
FEATURES Location/Qualifiers
promoter 11..422
/note="SV40 enhancerand early Pm"
/label=SV40 enhancer andearly Pm
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
intron 490..626
/label=intron
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
promoter 670..688
/note="T7 Pm"
/label=T7 Pm
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
gene 698..1630
/gene="Rluc"
/label=Rluc
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
CDS 698..1630
/gene="Rluc"
/codon_start=1
/transl_table=11
/product="Renillaluciferase"
/protein_id="AAC40213.1"
/db_xref="GI:3123920"
/translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAE
NAVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLT
AWFELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIE
EDIALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLS
WPREIPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNT
EFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
/label=Renilla luciferase
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
misc_feature 1633..1661
/note="polylinker"
/label=polylinker
/ApEinfo_fwdcolor=#e9e316
/ApEinfo_revcolor=#e9e316
gene 1663..3312
/gene="luc"
/label=luc
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
CDS 1663..3312
/gene="luc"
/codon_start=1
/transl_table=11
/product="fireflyluciferase"
/protein_id="AAC40214.1"
/db_xref="GI:3123921"
/translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
PANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGF
QSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRF
SHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSL
QDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPG
IRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVR
GPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAP
AELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTA
KKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
/label=firefly luciferase
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
polyA_signal 3363..3565
/note="SV40 latepolyadenylation signal"
/label=SV40 latepolyadenylation signal
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
gene 3719..4579
/gene="bla"
/label=bla
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
CDS 3719..4579
/gene="bla"
/function="ampicillinresistance"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="AAC40215.1"
/db_xref="GI:3123922"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
/label=beta-lactamase
/ApEinfo_fwdcolor=pink
/ApEinfo_revcolor=pink
ORIGIN
1 agatcgatct gcgcagcacc atggcctgaaataacctctg aaagaggaac ttggttaggt
61 accttctgag gcggaaagaa ccagctgtggaatgtgtgtc agttagggtg tggaaagtcc
121 ccaggctccc cagcaggcag aagtatgcaaagcatgcatc tcaattagtc agcaaccagg
181 tgtggaaagt ccccaggctc cccagcaggcagaagtatgc aaagcatgca tctcaattag
241 tcagcaacca tagtcccgcc cctaactccgcccatcccgc ccctaactcc gcccagttcc
301 gcccattctc cgccccatgg ctgactaattttttttattt atgcagaggc cgaggccgcc
361 tcggcctctg agctattcca gaagtagtgaggaggctttt ttggaggcct aggcttttgc
421 aaaaagcttg attcttctga cacaacagtctcgaacttaa gctgcagaag ttggtcgtga
481 ggcactgggc aggtaagtat caaggttacaagacaggttt aaggagacca atagaaactg
541 ggcttgtcga gacagagaag actcttgcgtttctgatagg cacctattgg tcttactgac
601 atccactttg cctttctctc cacaggtgtccactcccagt tcaattacag ctcttaaggc
661 tagagtactt aatacgactc actataggctagccaccatg acttcgaaag tttatgatcc
721 agaacaaagg aaacggatga taactggtccgcagtggtgg gccagatgta aacaaatgaa
781tgttcttgat tcatttatta attattatga ttcagaaaaa catgcagaaa atgctgttat
841 ttttttacat ggtaacgcgg cctcttcttatttatggcga catgttgtgc cacatattga
901 gccagtagcg cggtgtatta taccagaccttattggtatg ggcaaatcag gcaaatctgg
961 taatggttct tataggttac ttgatcattacaaatatctt actgcatggt ttgaacttct
1021 taatttacca aagaagatca tttttgtcggccatgattgg ggtgcttgtt tggcatttca
1081 ttatagctat gagcatcaag ataagatcaaagcaatagtt cacgctgaaa gtgtagtaga
1141 tgtgattgaa tcatgggatg aatggcctgatattgaagaa gatattgcgt tgatcaaatc
1201 tgaagaagga gaaaaaatgg ttttggagaataacttcttc gtggaaacca tgttgccatc
1261 aaaaatcatg agaaagttag aaccagaagaatttgcagca tatcttgaac cattcaaaga
1321 gaaaggtgaa gttcgtcgtc caacattatcatggcctcgt gaaatcccgt tagtaaaagg
1381 tggtaaacct gacgttgtac aaattgttaggaattataat gcttatctac gtgcaagtga
1441 tgatttacca aaaatgttta ttgaatcggacccaggattc ttttccaatg ctattgttga
1501 aggtgccaag aagtttccta atactgaatttgtcaaagta aaaggtcttc atttttcgca
1561 agaagatgca cctgatgaaa tgggaaaatatatcaaatcg ttcgttgagc gagttctcaa
1621 aaatgaacaa atgtcgacgg atccttcaacttccctgagc tcgaagacgc caaaaacata
1681 aagaaaggcc cggcgccatt ctatcctctagaggatggaa ccgctggaga gcaactgcat
1741 aaggctatga agagatacgc cctggttcctggaacaattg cttttacaga tgcacatatc
1801 gaggtgaaca tcacgtacgc ggaatacttcgaaatgtccg ttcggttggc agaagctatg
1861 aaacgatatg ggctgaatac aaatcacagaatcgtcgtat gcagtgaaaa ctctcttcaa
1921 ttctttatgc cggtgttggg cgcgttatttatcggagttg cagttgcgcc cgcgaacgac
1981 atttataatg aacgtgaatt gctcaacagtatgaacattt cgcagcctac cgtagtgttt
2041 gtttccaaaa aggggttgca aaaaattttgaacgtgcaaa aaaaattacc aataatccag
2101 aaaattatta tcatggattc taaaacggattaccagggat ttcagtcgat gtacacgttc
2161 gtcacatctc atctacctcc cggttttaat gaatacgattttgtaccaga gtcctttgat
2221 cgtgacaaaa caattgcact gataatgaattcctctggat ctactgggtt acctaagggt
2281 gtggcccttc cgcatagaac tgcctgcgtcagattctcgc atgccagaga tcctattttt
2341 ggcaatcaaa tcattccgga tactgcgattttaagtgttg ttccattcca tcacggtttt
2401 ggaatgttta ctacactcgg atatttgatatgtggatttc gagtcgtctt aatgtataga
2461 tttgaagaag agctgttttt acgatcccttcaggattaca aaattcaaag tgcgttgcta
2521 gtaccaaccc tattttcatt cttcgccaaaagcactctga ttgacaaata cgatttatct
2581 aatttacacg aaattgcttc tgggggcgcacctctttcga aagaagtcgg ggaagcggtt
2641 gcaaaacgct tccatcttcc agggatacgacaaggatatg ggctcactga gactacatca
2701 gctattctga ttacacccga gggggatgataaaccgggcg cggtcggtaa agttgttcca
2761 ttttttgaag cgaaggttgt ggatctggataccgggaaaa cgctgggcgt taatcagaga
2821 ggcgaattat gtgtcagagg acctatgattatgtccggtt atgtaaacaa tccggaagcg
2881 accaacgcct tgattgacaa ggatggatggctacattctg gagacatagc ttactgggac
2941 gaagacgaac acttcttcat agttgaccgcttgaagtctt taattaaata caaaggatat
3001 caggtggccc ccgctgaatt ggaatcgatattgttacaac accccaacat cttcgacgcg
3061 ggcgtggcag gtcttcccga cgatgacgccggtgaacttc ccgccgccgt tgttgttttg
3121 gagcacggaa agacgatgac ggaaaaagagatcgtggatt acgtcgccag tcaagtaaca
3181 accgcgaaaa agttgcgcgg aggagttgtgtttgtggacg aagtaccgaa aggtcttacc
3241 ggaaaactcg acgcaagaaa aatcagagagatcctcataa aggccaagaa gggcggaaag
3301 tccaaattgt aacacgtgta attctagagcggccgcttcg agcagacatg ataagataca
3361 ttgatgagtt tggacaaacc acaactagaatgcagtgaaa aaaatgcttt atttgtgaaa
3421 tttgtgatgc tattgcttta tttgtaaccattataagctg caataaacaa gttaacaaca
3481 acaattgcat tcattttatg tttcaggttcagggggaggt gtgggaggtt ttttaaagca
3541 agtaaaacct ctacaaatgt ggtaaaatcgataagtttaa acagatccag gtggcacttt
3601 tcggggaaat gtgcgcggaa cccctatttgtttatttttc taaatacatt caaatatgta
3661 tccgctcatg agacaataac cctgataaatgcttcaataa tattgaaaaa ggaagagtat
3721 gagtattcaa catttccgtg tcgcccttattccctttttt gcggcatttt gccttcctgt
3781 ttttgctcac ccagaaacgc tggtgaaagtaaaagatgct gaagatcagt tgggtgcacg
3841 agtgggttac atcgaactgg atctcaacagcggtaagatc cttgagagtt ttcgccccga
3901 agaacgtttt ccaatgatga gcacttttaaagttctgcta tgtggcgcgg tattatcccg
3961 tattgacgcc gggcaagagc aactcggtcgccgcatacac tattctcaga atgacttggt
4021 tgagtactca ccagtcacag aaaagcatcttacggatggc atgacagtaa gagaattatg
4081 cagtgctgcc ataaccatga gtgataacactgcggccaac ttacttctga caacgatcgg
4141 aggaccgaag gagctaaccg cttttttgcacaacatgggg gatcatgtaa ctcgccttga
4201 tcgttgggaa ccggagctga atgaagccataccaaacgac gagcgtgaca ccacgatgcc
4261 tgtagcaatg gcaacaacgt tgcgcaaactattaactggc gaactactta ctctagcttc
4321 ccggcaacaa ttaatagact ggatggaggcggataaagtt gcaggaccac ttctgcgctc
4381 ggcccttccg gctggctggt ttattgctgataaatctgga gccggtgagc gtgggtctcg
4441 cggtatcatt gcagcactgg ggccagatggtaagccctcc cgtatcgtag ttatctacac
4501 gacggggagt caggcaacta tggatgaacgaaatagacag atcgctgaga taggtgcctc
4561 actgattaag cattggtaac tgtcagaccaagtttactca tatatacttt agattgattt
4621aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
4681 caaaatccct taacgtgagt tttcgttccactgagcgtca gaccccgtag aaaagatcaa
4741 aggatcttct tgagatcctt tttttctgcgcgtaatctgc tgcttgcaaa caaaaaaacc
4801 accgctacca gcggtggttt gtttgccggatcaagagcta ccaactcttt ttccgaaggt
4861 aactggcttc agcagagcgc agataccaaatactgtcctt ctagtgtagc cgtagttagg
4921 ccaccacttc aagaactctg tagcaccgcctacatacctc gctctgctaa tcctgttacc
4981 agtggctgct gccagtggcg ataagtcgtgtcttaccggg ttggactcaa gacgatagtt
5041 accggataag gcgcagcggt cgggctgaacggggggttcg tgcacacagc ccagcttgga
5101 gcgaacgacc tacaccgaac tgagatacctacagcgtgag ctatgagaaa gcgccacgct
5161 tcccgaaggg agaaaggcgg acaggtatccggtaagcggc agggtcggaa caggagagcg
5221 cacgagggag cttccagggg gaaacgcctggtatctttat agtcctgtcg ggtttcgcca
5281 cctctgactt gagcgtcgat ttttgtgatgctcgtcaggg gggcggagcc tatggaaaaa
5341 cgccagcaac gcggcctttt tacggttcctggccttttgc tggccttttg ctcacatggc
5401 tcgac
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