pMCS-Green-Renilla-Luc

载体基本信息

载体质粒图谱和多克隆位点信息

载体简介

pMCS-Green Renilla Luc 载体简介The Thermo Scientific pMCS-Renilla Luc vector is a multiple cloning site plasmid to designed to accept a promoter sequence for study of gene regulation using the intracellular Renilla luciferase reporter.pMCS-Renilla Luc 载体特征: Intracellular Green Renilla luciferase gene, optimized for high expression 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 originThe Green Renilla Luc Vectors contain a gene cloned from the sea pansy, Renilla reniformis. The gene encodes a bioluminescent Green Renilla luciferase (36kDa). The pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential.These vectors are subject to a limited use label license.

载体序列

LOCUS Exported 5263 bp ds-DNA circular SYN 30-JUL-2016DEFINITION Intracellular green Renilla luciferase vector for cloning a transcriptional regulatory element.ACCESSION .VERSION .KEYWORDS pMCS-Green Renilla LucSOURCE synthetic DNA construct ORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 5263) AUTHORS Thermo Scientific (Pierce) TITLE Direct SubmissionFEATURES Location/Qualifiers source 1..5263 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" misc_feature 1..42 /note="MCS" /note="multiple cloning site" CDS 49..984 /codon_start=1 /product="green-emitting variant of Renilla luciferase" /note="Renilla Luciferase" /note="human codon-optimized" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGSALAFHYAYEHQDRIKAIVHMESVVDVIESWMGWPDIEEEL ALIKSEEGEKMVLENNFFVETLLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ" polyA_signal 1007..1118 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" promoter 1241..1570 /note="SV40 promoter" /note="SV40 enhancer and early promoter" rep_origin 1421..1556 /note="SV40 ori" /note="SV40 origin of replication" promoter 1618..1665 /note="EM7 promoter" /note="synthetic bacterial promoter " CDS 1684..2283 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /note="PuroR" /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" polyA_signal 2413..2534 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" CDS complement(2587..3447) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3711..4299 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 5041..5057 /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 5172..5263 /note="pause site" /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene"ORIGIN 1 actagtgaat tcggtaccga gctcctcgag aagcttggat ccgccaccat ggccagcaag 61 gtgtacgacc ccgagcagcg gaagcggatg atcaccggcc ctcagtggtg ggctcggtgc 121 aagcagatga acgtgctgga cagcttcatc aactactacg acagcgagaa gcacgccgag 181 aacgccgtga tcttcctgca cggcaacgcc accagcagct acctgtggcg gcacgtggtg 241 ccccacatcg agcctgtggc cagatgcatc atccccgacc tgatcggcat gggcaagagc 301 ggcaagtccg gcaacggcag ctaccggctg ctggaccact acaagtacct gaccgcttgg 361 tttgagctgc tgaacctgcc caagaagatc atcttcgtcg gccacgactg gggcagcgcc 421 ctggcctttc actacgccta cgagcaccag gaccggatca aggccatcgt gcacatggaa 481 agcgtggtgg acgtgatcga gagctggatg ggctggcccg acatcgagga agaactggcc 541 ctgatcaaga gcgaagaggg cgagaagatg gtgctggaaa acaacttctt cgtggaaacc 601 ctgctgccca gcaagatcat gcggaagctg gaacccgaag agttcgccgc ctacctggaa 661 cccttcaaag aaaagggcga agtgcggagg cccaccctga gctggcccag agagatcccc 721 ctggtcaagg gcggcaagcc cgacgtggtg cagatcgtgc ggaactacaa cgcctacctg 781 cgggccagcg acgacctgcc taagctgttc atcgagagcg accccggctt cttcagcaac 841 gccatcgtgg aaggcgccaa gaagttcccc aacaccgagt tcgtgaaagt gaagggcctg 901 cacttcctcc aggaagatgc ccccgacgag atgggcaagt acatcaagag cttcgtggaa 961 cgggtgctga agaacgagca gtgagcggcc gcaaaatcag cctcgactgt gccttctagt 1021 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 1081 cccactgtcc tttcctaata aaatgaggaa attgcatcac aacactcaac cctatctcgg 1141 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 1201 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg 1261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 1321 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 1381 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 1441 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 1501 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 1561 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt 1621 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa 1681 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc 1741 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg 1801 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc 1861 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg 1921 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc 1981 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag 2041 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg 2101 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc 2161 ctggagacat ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc 2221 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc 2281 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 2341 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 2401 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 2461 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 2521 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 2581 tggtcattac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 2641 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 2701 tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg atttatcagc 2761 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 2821 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 2881 gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 2941 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 3001 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 3061 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 3121 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 3181 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 3241 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 3301 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 3361 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 3421 ggcgacacgg aaatgttgaa tactcatatt cttccttttt caatattatt gaagcattta 3481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 3541 aggggtcagt gttacaacca attaaccaat tctgaacatt atcgcgagcc catttatacc 3601 tgaatatggc tcataacacc ccttgctcat gaccaaaatc ccttaacgtg agttacgcgc 3661 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 3721 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 3781 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 3841 cagataccaa atactgttct tctagtgtag ccgtagttag cccaccactt caagaactct 3901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 3961 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 4021 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 4081 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 4141 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 4201 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 4261 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 4321 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct 4381 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga 4441 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag gcgagagtag ggaactgcca 4501 ggcatcaaac taagcagaag gcccctgacg gatggccttt ttgcgtttct acaaactctt 4561 tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt tctgataacg 4621 agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg 4681 agtttaggga aagagcattt gtcagaatat ttaagggcgc ctgtcacttt gcttgatata 4741 tgagaattat ttaaccttat aaatgagaaa aaagcaacgc actttaaata agatacgttg 4801 ctttttcgat tgatgaacac ctataattaa actattcatc tattatttat gattttttgt 4861 atatacaata tttctagttt gttaaagaga attaagaaaa taaatctcga aaataataaa 4921 gggaaaatca gtttttgata tcaaaattat acatgtcaac gataatacaa aatataatac 4981 aaactataag atgttatcag tatttattat catttagaat aaattttgtg tcgcccttaa 5041 ttgtgagcgg ataacaatta cgagcttcat gcacagtggc gttgacattg attattgact 5101 agcatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat 5161 tagttattaa taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa 5221 taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tct