- 规格:20μl质粒
- 复制子:pUC
- 原核抗性:氨苄青霉素Amp
- 筛选标记:嘌呤霉素Puro
- 克隆菌株:大肠杆菌DH5α
- 质粒简介: pGL4.22[luc2CP/Puro]是一个哺乳细胞单荧光素酶报告质粒。The pGL4.22[luc2CP/Puro] Vector encodes the luciferase reporter gene luc2CP (Photinus pyralis) and is designed for high expression and reduced anomalous transcription. This vector 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. The vector is engineered with fewer consensus regulatory sequences for reduced background and a decreased risk of anomolous transcription and has a synthetic reporter gene, which has been codon optimized for mammalian expression. The pGL4.22[luc2CP/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 luc2CP reporter gene contains two protein destabilization sequences, hCL1 and hPEST. The protein encoded by luc2CP responds more quickly and with greater magnitude to changes in transcriptional activity than the luc2 gene, its more stable counterpart.
- 质粒图谱1: png/1-1P106110035T9.png
质粒序列: 点击查看序列LOCUS Exported 5583 bp ds-DNA circular SYN 06-JAN-2018DEFINITION synthetic circular DNAFEATURES Location/Qualifierssource 1..5583/organism="synthetic DNA construct"/mol_type="other DNA"regulatory 94..103/regulatory_class="other"/note="vertebrate consensus sequence for strong initiationof translation (Kozak, 1987)"CDS 100..1749/codon_start=1/product="firefly luciferase"/label=luciferase/note="synthetic luc2 version of the luciferase gene"/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAHIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"CDS 1756..1803/codon_start=1/product="non-ORF yeast peptide conferringubiquitin-dependent degradation"/label=hCL1/note="human codon-optimized"/translation="ACKNWFSSLSHFVIHL"CDS 1807..1926/codon_start=1/product="PEST degradation sequence from mouse ornithinedecarboxylase"/label=hPEST/note="human codon-optimized"/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"polyA_signal 1975..2096/label=SV40 poly(A) signal/note="SV40 polyadenylation signal"promoter 2290..2647/label=SV40 promoter/note="SV40 enhancer and early promoter"rep_origin 2498..2633/label=SV40 ori/note="SV40 origin of replication"CDS 2678..3277/codon_start=1/product="puromycin N-acetyltransferase"/label=PuroR/note="confers resistance to puromycin"/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"polyA_signal 3302..3350/note="synthetic polyadenylation signal"rep_origin complement(3677..4265)/direction=LEFT/label=ori/note="high-copy-number ColE1/pMB1/pBR322/pUC origin ofreplication"CDS complement(4465..5325)/codon_start=1/product="beta-lactamase"/label=AmpR/note="confers resistance to ampicillin, carbenicillin, andrelated antibiotics"/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"polyA_signal 5430..5478/note="synthetic polyadenylation signal"misc_feature 5492..5583/label=pause site/note="RNA polymerase II transcriptional pause signal fromthe human alpha-2 globin gene"ORIGIN1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaagatc tggcctcggc61 ggccaagctt ggcaatccgg tactgttggt aaagccacca tggaagatgc caaaaacatt121 aagaagggcc cagcgccatt ctacccactc gaagacggga ccgccggcga gcagctgcac181 aaagccatga agcgctacgc cctggtgccc ggcaccatcg cctttaccga cgcacatatc241 gaggtggaca ttacctacgc cgagtacttc gagatgagcg ttcggctggc agaagctatg301 aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag361 ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg ctgtggcccc agctaacgac421 atctacaacg agcgcgagct gctgaacagc atgggcatca gccagcccac cgtcgtattc481 gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa agaagctacc gatcatacaa541 aagatcatca tcatggatag caagaccgac taccagggct tccaaagcat gtacaccttc601 gtgacttccc atttgccacc cggcttcaac gagtacgact tcgtgcccga gagcttcgac661 cgggacaaaa ccatcgccct gatcatgaac agtagtggca gtaccggatt gcccaagggc721 gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc781 ggcaaccaga tcatccccga caccgctatc ctcagcgtgg tgccatttca ccacggcttc841 ggcatgttca ccacgctggg ctacttgatc tgcggctttc gggtcgtgct catgtaccgc901 ttcgaggagg agctattctt gcgcagcttg caagactata agattcaatc tgccctgctg961 gtgcccacac tatttagctt cttcgctaag agcactctca tcgacaagta cgacctaagc1021 aacttgcacg agatcgccag cggcggggcg ccgctcagca aggaggtagg tgaggccgtg1081 gccaaacgct tccacctacc aggcatccgc cagggctacg gcctgacaga aacaaccagc1141 gccattctga tcacccccga aggggacgac aagcctggcg cagtaggcaa ggtggtgccc1201 ttcttcgagg ctaaggtggt ggacttggac accggtaaga cactgggtgt gaaccagcgc1261 ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct acgttaacaa ccccgaggct1321 acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg gcgacatcgc ctactgggac1381 gaggacgagc acttcttcat cgtggaccgg ctgaagagcc tgatcaaata caagggctac1441 caggtagccc cagccgaact ggagagcatc ctgctgcaac accccaacat cttcgacgcc1501 ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg1561 gaacacggta aaaccatgac cgagaaggag atcgtggact atgtggccag ccaggttaca1621 accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc1681 ggcaagttgg acgcccgcaa gatccgcgag attctcatta aggccaagaa gggcggcaag1741 atcgccgtga attctgcttg caagaactgg ttcagtagct taagccactt tgtgatccac1801 cttaacagcc acggcttccc tcccgaggtg gaggagcagg ccgccggcac cctgcccatg1861 agctgcgccc aggagagcgg catggataga caccctgctg cttgcgccag cgccaggatc1921 aacgtctaac tgcagtctag agtcggggcg gccggccgct tcgagcagac atgataagat1981 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg2041 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca2101 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa2161 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtttgcg tattgggcgc2221 tcttccgctg atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt2281 agctaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa2341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac2401 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa2461 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag2521 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc2581 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt2641 ttgcaaaaag ctcgattctt ctgacactag cgccaccatg accgagtaca agcctaccgt2701 gcgcctggcc actcgcgatg atgtgccccg cgccgtccgc actctggccg ccgctttcgc2761 cgactacccc gctacccggc acaccgtgga ccccgaccgg cacatcgagc gtgtgacaga2821 gttgcaggag ctgttcctga cccgcgtcgg gctggacatc ggcaaggtgt gggtagccga2881 cgacggcgcg gccgtggccg tgtggactac ccccgagagc gttgaggccg gcgccgtgtt2941 cgccgagatc ggcccccgaa tggccgagct gagcggcagc cgcctggccg cccagcagca3001 aatggagggc ctgcttgccc cccatcgtcc caaggagcct gcctggtttc tggccactgt3061 aggagtgagc cccgaccacc agggcaaggg cttgggcagc gccgtcgtgt tgcccggcgt3121 agaggccgcc gaacgcgccg gtgtgcccgc ctttctcgaa acaagcgcac caagaaacct3181 tccattctac gagcgcctgg gcttcaccgt gaccgccgat gtcgaggtgc ccgagggacc3241 taggacctgg tgtatgacac gaaaacctgg cgcctaatga tctagaaccg gtcatggccg3301 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg ttcgaactag3361 atgctgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg3421 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca3481 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc3541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg3781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct3841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg3901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct3961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac4021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt4081 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc4141 tgctgaagcc agttaccttc ggaaaaagag![]()