pCMV-Gaussia-Dura Luc 载体简介The Thermo Scientific pCMV-Gaussia-Dura Luc vector is a derivative of pMCS-Gaussia-Dura Luc. It contains a mutant form of the Gaussia luciferase gene (conferring better bioluminescent signal stability than the native luciferase) under the control of the strong Cytomegalovirus (CMV) promoter. This constitutive expression vector can be used as a normalization control to account for experimental variation in combination with other reporters.载体特征: Strong (CMV) constitutive promoter for co-transfection and normalization Naturally-secreting Gaussia-Dura luciferase gene, optimized for high expression and glow-stability in mammalian systems Multiple cloning site (MCS) provides versatility for transfer of regulatory elements from one plasmid to another Transcription termination site (Ter), Lac operator (Lac O1), and transcriptional pause site (TPS) used to minimize background by reducing transcriptional read-through Both puromycin (Pur) and ampicillin (Amp) markers for drug selection in mammalian and bacterial cells, respectively High-copy pUC bacterial DNA replication originGaussia-Dura luciferase (approx. 20kDa) is a secreted protein that enables measurement of the reporter activity in media (for real-time assays) and in cell lysates. The parent pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential.
LOCUS Exported 5482 bp ds-DNA circular SYN 24-JUL-2016DEFINITION Control vector for constitutive high-level expression of secreted Gaussia-Dura luciferase under control of the CMV promoter.ACCESSION .VERSION .KEYWORDS pCMV-Gaussia-Dura LucSOURCE synthetic DNA construct ORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 5482) AUTHORS Thermo Scientific (Pierce) TITLE Direct Submission JOURNAL Exported Tuesday, July 26, 2016 from SnapGene Viewer 3.1.4 http://www.snapgene.comFEATURES Location/Qualifiers source 1..5482 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" enhancer 63..366 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 367..570 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" promoter 615..633 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 646..1203 /codon_start=1 /product="enhanced variant of secreted Gaussia luciferase" /note="Gaussia-Dura Luc" /note="human codon-optimized" /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG KLPGKKLPLEVLKELEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA QGGIGEAIVDIPEIPGFKDLEPLEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ RCATFASKIQGQVDKIKGAGGD" polyA_signal 1226..1337 /note="BGH pA" /note="bovine growth hormone polyadenylation signal" promoter 1460..1789 /note="SV40 promoter" /note="SV40 enhancer and early promoter" rep_origin 1640..1775 /note="SV40 ori" /note="SV40 origin of replication" promoter 1837..1884 /note="EM7 promoter" /note="synthetic bacterial promoter " CDS 1903..2502 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /note="Puromycin" /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" polyA_signal 2632..2753 /note="SV40 pA" /note="SV40 polyadenylation signal" CDS complement(2806..3666) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="Ampicillin" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3930..4518 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 5260..5276 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." misc_feature 5391..5482 /note="TPS" /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene"ORIGIN 1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc 61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg 421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt 541 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac 601 tggcttatcg aaattaatac gactcactat aggggatccg ccaccatggg agtcaaagtt 661 ctgtttgccc tgatctgcat cgctgtggcc gaggccaagc ccaccgagaa caacgaagac 721 ttcaacatcg tggccgtggc cagcaacttc gcgaccacgg atctcgatgc tgaccgcggg 781 aagttgcccg gcaagaagct gccgctggag gtgctcaaag agttggaagc caatgcccgg 841 aaagctggct gcaccagggg ctgtctgatc tgcctgtccc acatcaagtg cacgcccaag 901 atgaagaagt tcatcccagg acgctgccac acctacgaag gcgacaaaga gtccgcacag 961 ggcggcatag gcgaggcgat cgtcgacatt cctgagattc ctgggttcaa ggacttggag 1021 cccttggagc agttcatcgc acaggtcgat ctgtgtgtgg actgcacaac tggctgcctc 1081 aaagggcttg ccaacgtgca gtgttctgac ctgctcaaga agtggctgcc gcaacgctgt 1141 gcgacctttg ccagcaagat ccagggccag gtggacaaga tcaagggggc cggtggtgac 1201 taagcggccg caaaatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc 1261 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa 1321 aatgaggaaa ttgcatcaca acactcaacc ctatctcggt ctattctttt gatttataag 1381 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 1441 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 1501 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 1561 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 1621 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 1681 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 1741 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 1801 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag 1861 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatgaccga gtacaagccc 1921 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 1981 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 2041 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 2101 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 2161 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 2221 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 2281 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 2341 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacatc cgcgccccgc 2401 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 2461 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacacgtgct acgagatttc 2521 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 2581 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 2641 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 2701 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 2761 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcattacc aatgcttaat 2821 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 2881 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagcg ctgcgatgat 2941 accgcgagaa ccacgctcac cggctccgga tttatcagca ataaaccagc cagccggaag 3001 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 3061 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccatcgc 3121 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 3181 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 3241 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 3301 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 3361 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 3421 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 3481 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 3541 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 3601 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 3661 actcatattc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 3721 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa 3781 ttaaccaatt ctgaacatta tcgcgagccc atttatacct gaatatggct cataacaccc 3841 cttgctcatg accaaaatcc cttaacgtga gttacgcgcg cgtcgttcca ctgagcgtca 3901 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 3961 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 4021 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 4081 ctagtgtagc cgtagttagc ccaccacttc aagaactctg tagcaccgcc tacatacctc 4141 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 4201 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 4261 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 4321 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 4381 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 4441 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 4501 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 4561 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 4621 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 4681 gtgagcgagg aagcggaagg cgagagtagg gaactgccag gcatcaaact aagcagaagg 4741 cccctgacgg atggcctttt tgcgtttcta caaactcttt ctgtgttgta aaacgacggc 4801 cagtcttaag ctcgggcccc ctgggcggtt ctgataacga gtaatcgtta atccgcaaat 4861 aacgtaaaaa cccgcttcgg cgggtttttt tatgggggga gtttagggaa agagcatttg 4921 tcagaatatt taagggcgcc tgtcactttg cttgatatat gagaattatt taaccttata 4981 aatgagaaaa aagcaacgca ctttaaataa gatacgttgc tttttcgatt gatgaacacc 5041 tataattaaa ctattcatct attatttatg attttttgta tatacaatat ttctagtttg 5101 ttaaagagaa ttaagaaaat aaatctcgaa aataataaag ggaaaatcag tttttgatat 5161 caaaattata catgtcaacg ataatacaaa atataataca aactataaga tgttatcagt 5221 atttattatc atttagaata aattttgtgt cgcccttaat tgtgagcgga taacaattac 5281 gagcttcatg cacagtggcg ttgacattga ttattgacta gcatgttctt tcctgcgtta 5341 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat aacatacgct 5401 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt 5461 gcaggtgcca gaacatttct ct