pMCS-Red-Firefly-Luc载体简介
Thermo Scientific pMCS-Red Firefly Luc is a multiple cloning site plasmid designed to accept a promoter sequence for the study of gene regulation using the intracellular red firefly luciferase reporter with excellent light intensity..
Features of pMCS-Red Firefly Luc:
• Intracellular Red Firefly 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
• Two control vectors available with strong (CMV) or weak (TK) constitutive promoters for co-transfection and normalization
Features of Red Firefly Luc:
• Red Firefly Luc Vectors contains a mutant form of the firefly luciferase gene that has a red-shifted emission spectrum.
• The pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential.
• The pCMV and pTK vectors have the luciferase gene under the CMV (Cytomegalovirus) promoter and Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter, respectively.
• These pCMV and pTK constitutive expression vectors can be used as normalization controls to account for experimental variation in combination with other reporters.
LOCUS Exported File 5977 bp ds-DNA circular SYN 24-七月-2016
DEFINITION Intracellular red firefly luciferase vector for cloning a
transcriptional regulatory element.
ACCESSION .
VERSION .
KEYWORDS pMCS-Red Firefly Luc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5977)
AUTHORS Thermo Scientific (Pierce)
TITLE Direct Submission
JOURNAL Exported 2016-07-24 from SnapGene Viewer 1.5.3
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5977
/organism="synthetic DNA construct"
/lab_host="Mammalian Cells"
/mol_type="other DNA"
misc_feature 1..42
/note="MCS"
/note="multiple cloning site"
/note="color: #00ff00"
CDS 49..1695
/codon_start=1
/product="intracellular red firefly luciferase"
/note="Red Firefly Luciferase"
/note="human codon-optimized"
/note="color: #ff0000"
/translation="MENMENDENIVVGPKPFYPIEEGSAGTQLRKYMERYAKLGAIAFT
NAVTGVDYSYAEYLEKSCCLGKALQNYGLVVDGRIALCSENCEEFFIPVIAGLFIGVGV
APTNEIYTLRELVHSLGISKPTIVFSSKKGLDKVITVQKTVTTIKTIVILDSKVDYRGY
QCLDTFIKRNTPPGFQASSFKTVEVDRKEQVALIMNSSGSTGLPKGVQLTHENTVTRFS
HARDPIYGNQVSPGTAVLTVVPFHHGFGMFTTLGYLICGFRVVMLTKFDEETFLKTLQD
YKCTYVILVPTLFAILNKSELLNKYDLSNLVEIASGGAPLSKEVGEAVARRFNLPGVRQ
GYGLTETTSAIIITPEGDDKPGASGKVVPLFKAKVIDLDTKKSLGPNRRGEVCVKGPML
MKGYVNNPEATKELIDEEGWLHTGDIGYYDEEKHFFIVDRLKSLIKYKGYQVPPAELES
VLLQHPSIFDAGVAGVPDPVAGELPGAVVVLESGKNMTEKEVMDYVASQVSNAKRLRGG
VRFVDEVPKGLTGKIDGRAIREILKKPVAKM"
polyA_signal 1707..1832
/note="BGH pA"
/note="bovine growth hormone polyadenylation signal
bGH poly(A) signal"
/note="color: #ffffff"
promoter 1955..2284
/note="pSV40"
/note="SV40 enhancer and early promoter"
/note="color: #000000; direction: RIGHT"
rep_origin 2135..2270
/note="SV40 ori"
/note="SV40 origin of replication"
/note="color: #ffff00"
promoter 2332..2379
/note="EM7 promoter"
/note="synthetic bacterial promoter?"
/note="color: #000000; direction: RIGHT"
CDS 2398..2997
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="Puromycin"
/note="confers resistance to puromycin"
/note="color: #33cccc"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 3127..3248
/note="SV40 pA"
/note="SV40 polyadenylation signal"
/note="color: #ffffff"
CDS complement(3301..4161)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/note="This feature has 2 segments:
???1:?3301?..?4092?/?#ccffcc
???2:?4093?..?4161?/?#ccffcc?/?signal?sequence
Cleavage site after base 4092"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4425..5013
/direction=RIGHT
/note="pUC ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/note="color: #ffff00"
misc_feature 5145..5752
/note="Terminator"
/note="Transcriptional terminator"
/note="color: #808080"
protein_bind 5755..5771
/bound_moiety="lac repressor encoded by lacI"
/note="lac O1"
/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)."
/note="color: #31849b"
misc_feature 5886..5977
/note="TPS"
/note="RNA polymerase II transcriptional pause signal from
the human?alpha-2 globin gene
Transcriptional pause site"
/note="color: #99cc00"
ORIGIN
1 actagtgaat tcggtaccga gctcctcgag aagcttggat ccgccaccat ggaaaatatg
61 gaaaacgacg agaacatcgt ggtgggcccc aagcccttct accccatcga ggaaggcagc
121 gccggcaccc agctgcggaa gtacatggaa agatacgcca agctgggcgc cattgccttc
181 accaacgccg tgaccggcgt ggactacagc tacgccgagt acctggaaaa gagctgctgc
241 ctgggcaagg ctctgcagaa ctacggcctg gtggtggacg gccggatcgc cctgtgcagc
301 gagaactgcg aggaattctt catccccgtg atcgccggcc tgttcatcgg cgtgggcgtg
361 gctcccacca acgagatcta caccctgcgg gagctggtgc acagcctggg catcagcaag
421 cccaccatcg tgttcagcag caagaagggc ctggacaaag tcatcaccgt gcagaaaacc
481 gtgaccacca tcaagaccat cgtgatcctg gacagcaagg tggactaccg gggctaccag
541 tgcctggaca ccttcatcaa gcggaacacc ccccctggct tccaggccag cagcttcaag
601 accgtggagg tggaccggaa agaacaggtg gccctgatca tgaacagcag cggcagcacc
661 ggcctgccca agggcgtgca gctgacccac gagaacaccg tgacccggtt cagccacgcc
721 agggacccca tctacggcaa ccaggtgtcc cccggcaccg ccgtgctgac cgtggtgccc
781 ttccaccacg gcttcggcat gttcaccacc ctgggctacc tgatctgcgg cttccgggtg
841 gtgatgctga ccaagttcga cgaggaaacc ttcctgaaaa ccctgcagga ctacaagtgc
901 acctacgtga ttctggtgcc caccctgttc gccatcctga acaagagcga gctgctgaac
961 aagtacgacc tgagcaacct ggtggagatc gccagcggcg gagcccccct gagcaaagaa
1021 gtgggagagg ccgtcgccag gcggttcaat ctgcccggcg tgcggcaggg ctacggcctg
1081 accgagacaa ccagcgccat catcatcacc cccgagggcg acgacaagcc tggagccagc
1141 ggcaaggtgg tgcccctgtt caaggccaaa gtgatcgacc tggacaccaa gaagagcctg
1201 ggccccaaca gacggggcga agtgtgcgtg aagggcccca tgctgatgaa gggctacgtg
1261 aacaaccccg aggccaccaa agagctgatc gacgaagagg gctggctgca caccggcgac
1321 atcggctact acgacgaaga gaagcacttc ttcatcgtgg accggctgaa gagcctgatc
1381 aagtacaagg gctatcaggt gccccctgcc gagctggaaa gcgtcctgct gcagcacccc
1441 agcatcttcg acgccggcgt ggccggggtg ccagatcctg tggccggcga gctgcctggc
1501 gccgtggtgg tgctggaatc cggcaagaac atgaccgaga aagaagtgat ggactacgtc
1561 gccagccagg tgtccaacgc caagcggctg agaggcggcg tgagattcgt ggacgaagtg
1621 ccaaagggcc tgaccggcaa gatcgacggc agggccatcc gggagatcct gaagaaaccc
1681 gtggccaaga tgtgatgagc ggccgcaaaa tcagcctcga ctgtgccttc tagttgccag
1741 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
1801 gtcctttcct aataaaatga ggaaattgca tcacaacact caaccctatc tcggtctatt
1861 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
1921 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
1981 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
2041 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
2101 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
2161 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
2221 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
2281 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
2341 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
2401 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
2461 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
2521 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
2581 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
2641 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
2701 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
2761 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
2821 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
2881 acatccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
2941 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac
3001 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
3061 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
3121 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
3181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
3241 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
3301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
3361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
3421 cagcgctgcg atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa
3481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
3541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
3601 cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
3661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
3721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
3781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
3841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
3901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
3961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
4021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
4081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
4141 acggaaatgt tgaatactca tattcttcct ttttcaatat tattgaagca tttatcaggg
4201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
4261 cagtgttaca accaattaac caattctgaa cattatcgcg agcccattta tacctgaata
4321 tggctcataa caccccttgc tcatgaccaa aatcccttaa cgtgagttac gcgcgcgtcg
4381 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
4441 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
4501 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
4561 ccaaatactg ttcttctagt gtagccgtag ttagcccacc acttcaagaa ctctgtagca
4621 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
4681 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
4741 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
4801 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
4861 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
4921 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
4981 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
5041 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
5101 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
5161 gagcgcagcg agtcagtgag cgaggaagcg gaaggcgaga gtagggaact gccaggcatc
5221 aaactaagca gaaggcccct gacggatggc ctttttgcgt ttctacaaac tctttctgtg
5281 ttgtaaaacg acggccagtc ttaagctcgg gccccctggg cggttctgat aacgagtaat
5341 cgttaatccg caaataacgt aaaaacccgc ttcggcgggt ttttttatgg ggggagttta
5401 gggaaagagc atttgtcaga atatttaagg gcgcctgtca ctttgcttga tatatgagaa
5461 ttatttaacc ttataaatga gaaaaaagca acgcacttta aataagatac gttgcttttt
5521 cgattgatga acacctataa ttaaactatt catctattat ttatgatttt ttgtatatac
5581 aatatttcta gtttgttaaa gagaattaag aaaataaatc tcgaaaataa taaagggaaa
5641 atcagttttt gatatcaaaa ttatacatgt caacgataat acaaaatata atacaaacta
5701 taagatgtta tcagtattta ttatcattta gaataaattt tgtgtcgccc ttaattgtga
5761 gcggataaca attacgagct tcatgcacag tggcgttgac attgattatt gactagcatg
5821 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
5881 ttaataacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
5941 gtccccagtg caagtgcagg tgccagaaca tttctct
//