pMCS-Red-Firefly-Luc

pMCS-Red-Firefly-Luc
 

载体基本信息

出品公司:	Ybscience
载体名称:	pMCS-Red-Firefly-Luc ; pMCS-Red Firefly Luc
质粒类型:	无启动子载体；哺乳动物载体；红色荧光素酶报告载体
高拷贝/低拷贝:	--
克隆方法:	限制性内切酶，多克隆位点
启动子:	无
载体大小:	5977 bp
5' 测序引物及序列:	--
3' 测序引物及序列:	--
载体标签:	Red Firefly Luciferase红色萤火虫荧光素酶
载体抗性:	氨苄青霉素
筛选标记:	嘌呤霉素（Puromycin）
克隆菌株:	TOP10等常规菌株
宿主细胞（系）:	常规哺乳动物细胞
备注:	--
稳定性:	稳表达
组成型/诱导型:	组成型
病毒/非病毒:	非病毒

 

载体质粒图谱和多克隆位点信息

载体简介

 

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.

 

载体序列

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