LOCUS       sequence-290892-        6043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of 6xbsMS2-Malat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6043)
  AUTHORS   Kuzuoglu-Ozturk D, Bhandari D, Huntzinger E, Fauser M, Helms S, 
            Izaurralde E
  TITLE     miRISC and the CCR4-NOT complex silence mRNA targets independently 
            of 43S ribosomal scanning.
  JOURNAL   EMBO J. 2016 Jun 1;35(11):1186-203. doi: 10.15252/embj.201592901. 
            Epub 2016 Mar 23.
  PUBMED    27009120
REFERENCE   2  (bases 1 to 6043)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.15252/embj.201592901"; journalName: "EMBO J"; date: "2016-06-1- 
            1"; volume: "35"; issue: "11"; pages: "1186-203"
COMMENT     Sequence Label: pRL-TK-hFluc-HCV-hRluc-6xbsMS2-Malat_AC
FEATURES             Location/Qualifiers
     source          1..6043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..759
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             766..2418
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(832..850)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2858..3793
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_RNA        3810..3828
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3849..3867
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3888..3906
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3927..3945
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3966..3984
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4005..4023
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        4252..4356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4357..4425,4426..5217)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4575..4594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5388..5976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5877..5896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct
       61 gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt
      121 cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc
      181 cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc
      241 gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg
      301 gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg
      361 gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac
      421 gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat
      481 cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat
      541 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
      601 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
      661 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
      721 ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttatgga agacgccaaa
      781 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
      841 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
      901 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
      961 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     1021 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     1081 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
     1141 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     1201 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     1261 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     1321 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct
     1381 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     1441 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     1501 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     1561 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     1621 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     1681 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     1741 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     1801 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     1861 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     1921 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     1981 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     2041 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     2101 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     2161 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     2221 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     2281 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     2341 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     2401 ggaaagatcg ccgtgtaatt ctagtgtacg tagccagccc ccgattgggg gcgacactcc
     2461 accatagatc actcccctgt gaggaactac tgtcttcacg cagaaagcgt ctagccatgg
     2521 cgttagtatg agagtcgtgc agcctccagg accccccctc ccgggagagc catagtggtc
     2581 tgcggaaccg gtgagtacac cggaattgcc aggacgaccg ggtcctttct tggatcaacc
     2641 cgctcaatgc ctggagattt gggcgtgccc ccgcaagact gctagccgag tagtgttggg
     2701 tcgcgaaagg ccttgtggta ctgcctgata gggtgcttgc gagtgccccg ggaggtctcg
     2761 tagaccgtgc accatgagca cgaatcctaa acctcaaaga aaaaccaaac gtaacaccaa
     2821 ccgccgccca caggacgtct tcatatgtct agccaccatg acttcgaaag tttatgatcc
     2881 agaacaaagg aaacggatga taactggtcc gcagtggtgg gccagatgta aacaaatgaa
     2941 tgttcttgat tcatttatta attattatga ttcagaaaaa catgcagaaa atgctgttat
     3001 ttttttacat ggtaacgcgg cctcttctta tttatggcga catgttgtgc cacatattga
     3061 gccagtagcg cggtgtatta taccagacct tattggtatg ggcaaatcag gcaaatctgg
     3121 taatggttct tataggttac ttgatcatta caaatatctt actgcatggt ttgaacttct
     3181 taatttacca aagaagatca tttttgtcgg ccatgattgg ggtgcttgtt tggcatttca
     3241 ttatagctat gagcatcaag ataagatcaa agcaatagtt cacgctgaaa gtgtagtaga
     3301 tgtgattgaa tcatgggatg aatggcctga tattgaagaa gatattgcgt tgatcaaatc
     3361 tgaagaagga gaaaaaatgg ttttggagaa taacttcttc gtggaaacca tgttgccatc
     3421 aaaaatcatg agaaagttag aaccagaaga atttgcagca tatcttgaac cattcaaaga
     3481 gaaaggtgaa gttcgtcgtc caacattatc atggcctcgt gaaatcccgt tagtaaaagg
     3541 tggtaaacct gacgttgtac aaattgttag gaattataat gcttatctac gtgcaagtga
     3601 tgatttacca aaaatgttta ttgaatcgga cccaggattc ttttccaatg ctattgttga
     3661 aggtgccaag aagtttccta atactgaatt tgtcaaagta aaaggtcttc atttttcgca
     3721 agaagatgca cctgatgaaa tgggaaaata tatcaaatcg ttcgttgagc gagttctcaa
     3781 aaatgaacaa taattctaga ctgtagaaaa catgaggatc acccatgtct gctggacgac
     3841 tgtagaaaac atgaggatca cccatgtctg ctgtctagct gtagaaaaca tgaggatcac
     3901 ccatgtctgc tggacgactg tagaaaacat gaggatcacc catgtctgct gtctagctgt
     3961 agaaaacatg aggatcaccc atgtctgctg gacgactgta gaaaacatga ggatcaccca
     4021 tgtctgctgg gccctagagc ggccgcgatt cgtcagtagg gttgtaaagg tttttctttt
     4081 cctgagaaaa caaccttttg ttttctcagg ttttgctttt tggcctttcc ctagctttaa
     4141 aaaaaaaaaa gcaaaagacg ctggtggctg gcactcctgg tttccaggac ggggttcaag
     4201 tccctgcggt gtctttgctt ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc
     4261 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4321 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4381 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4441 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4501 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4561 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4621 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4681 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4741 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4801 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4861 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4921 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4981 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5041 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5101 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5161 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5221 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5281 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5341 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5401 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5461 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5521 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5581 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5641 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5701 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5761 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5821 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5881 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5941 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6001 cggttcctgg ccttttgctg gccttttgct cacatggctc gac
//
