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 origin
The 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.
LOCUS Exported 5263 bp ds-DNA circular SYN 30-JUL-2016
DEFINITION Intracellular green Renilla luciferase vector for cloning a
transcriptional regulatory element.
ACCESSION .
VERSION .
KEYWORDS pMCS-Green Renilla Luc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5263)
AUTHORS Thermo Scientific (Pierce)
TITLE Direct Submission
FEATURES 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
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