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载体简介
pCMV-Green Renilla Luc 载体简介
The Thermo Scientific pCMV-Green Renilla Luc vector is a derivative of pMCS-Green Renilla Luc. It contains the luciferase gene under the control of the CMV (Cytomegalovirus) promoter, creating a constitutive expression vector that can be used as a normalization control to account for experimental variation in combination with other reporters.
pCMV-Green Renilla Luc 载体特征:
Strong (CMV) constitutive promoter for co-transfection and normalization
Intracellular Green Renilla luciferase gene, optimized for high expression in mammalian systems
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
载体序列
LOCUS Exported 5860 bp ds-DNA circular SYN 26-JUL-2013
DEFINITION Control vector for constitutive high-level expression of
intracellular green Renilla luciferase under control of the CMV
promoter.
ACCESSION .
VERSION .
KEYWORDS pCMV-Green Renilla Luc
REFERENCE 1 (bases 1 to 5860)
AUTHORS Thermo Scientific (Pierce)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5860
/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..1581
/codon_start=1
/product="green-emitting variant of Renilla luciferase"
/note="Green Renilla Luciferase"
/note="human codon-optimized"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGSALAFHYAYEHQDRIKAIVHMESVVDVIESWMGWPDIEEEL
ALIKSEEGEKMVLENNFFVETLLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
polyA_signal 1604..1715
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
promoter 1838..2167
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2018..2153
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 2215..2262
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 2281..2880
/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 3010..3131
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(3184..4044)
/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 4308..4896
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 5638..5654
/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 5769..5860
/note="pause site"
/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 ccaccatggc cagcaaggtg
661 tacgaccccg agcagcggaa gcggatgatc accggccctc agtggtgggc tcggtgcaag
721 cagatgaacg tgctggacag cttcatcaac tactacgaca gcgagaagca cgccgagaac
781 gccgtgatct tcctgcacgg caacgccacc agcagctacc tgtggcggca cgtggtgccc
841 cacatcgagc ctgtggccag atgcatcatc cccgacctga tcggcatggg caagagcggc
901 aagtccggca acggcagcta ccggctgctg gaccactaca agtacctgac cgcttggttt
961 gagctgctga acctgcccaa gaagatcatc ttcgtcggcc acgactgggg cagcgccctg
1021 gcctttcact acgcctacga gcaccaggac cggatcaagg ccatcgtgca catggaaagc
1081 gtggtggacg tgatcgagag ctggatgggc tggcccgaca tcgaggaaga actggccctg
1141 atcaagagcg aagagggcga gaagatggtg ctggaaaaca acttcttcgt ggaaaccctg
1201 ctgcccagca agatcatgcg gaagctggaa cccgaagagt tcgccgccta cctggaaccc
1261 ttcaaagaaa agggcgaagt gcggaggccc accctgagct ggcccagaga gatccccctg
1321 gtcaagggcg gcaagcccga cgtggtgcag atcgtgcgga actacaacgc ctacctgcgg
1381 gccagcgacg acctgcctaa gctgttcatc gagagcgacc ccggcttctt cagcaacgcc
1441 atcgtggaag gcgccaagaa gttccccaac accgagttcg tgaaagtgaa gggcctgcac
1501 ttcctccagg aagatgcccc cgacgagatg ggcaagtaca tcaagagctt cgtggaacgg
1561 gtgctgaaga acgagcagtg agcggccgca aaatcagcct cgactgtgcc ttctagttgc
1621 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
1681 actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct atctcggtct
1741 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
1801 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
1861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
1921 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
1981 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
2041 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
2101 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
2161 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
2221 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
2281 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
2341 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
2401 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
2461 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
2521 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
2581 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
2641 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
2701 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
2761 gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
2821 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
2881 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
2941 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
3001 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
3061 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
3121 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
3181 tcattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
3241 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
3301 ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt tatcagcaat
3361 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
3421 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
3481 caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
3541 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
3601 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
3661 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
3721 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
3781 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
3841 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
3901 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
3961 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
4021 gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa gcatttatca
4081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
4141 ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat ttatacctga
4201 atatggctca taacacccct tgctcatgac caaaatccct taacgtgagt tacgcgcgcg
4261 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
4321 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
4381 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
4441 ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa gaactctgta
4501 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
4561 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
4621 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
4681 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
4741 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
4801 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
4861 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
4921 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
4981 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
5041 accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga actgccaggc
5101 atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca aactctttct
5161 gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct gataacgagt
5221 aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta tggggggagt
5281 ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct tgatatatga
5341 gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt
5401 tttcgattga tgaacaccta taattaaact attcatctat tatttatgat tttttgtata
5461 tacaatattt ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa taataaaggg
5521 aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat ataatacaaa
5581 ctataagatg ttatcagtat ttattatcat ttagaataaa ttttgtgtcg cccttaattg
5641 tgagcggata acaattacga gcttcatgca cagtggcgtt gacattgatt attgactagc
5701 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
5761 ttattaataa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
5821 gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//
