LOCUS       sequence-357771-        5865 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 5865)
  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 5865)
  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-CrPV-hRluc-6xbsMS2-Malat_AC
FEATURES             Location/Qualifiers
     source          1..5865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..752
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      782..791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             788..2440
                     /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(854..872)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2673..3608
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_RNA        3625..3643
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3664..3682
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3703..3721
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3742..3760
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3781..3799
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3820..3838
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        4067..4171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4172..4240,4241..5032)
                     /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(4390..4409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5203..5791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5692..5711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatgagtctt cggacctcgc gggggccgct taagcggtgg ttagggtttg tctgacgcgg
       61 ggggaggggg aaggaacgaa acactctcat tcggaggcgg ctcggggttt ggtcttggtg
      121 gccacgggca cgcagaagag cgccgcgatc ctcttaagca cccccccgcc ctccgtggag
      181 gcgggggttt ggtcggcggg tggtaactgg cgggccgctg actcgggcgg gtcgcgcgcc
      241 ccagagtgtg accttttcgg tctgctcgca gacccccggg cggcgccgcc gcggcggcga
      301 cgggctcgct gggtcctagg ctccatgggg accgtatacg tggacaggct ctggagcatc
      361 cgcacgactg cggtgatatt accggagacc ttctgcggga cgagccgggt cacgcggctg
      421 acgcggagcg tccgttgggc gacaaacacc aggacggggc acaggtacac tatcttgtca
      481 cccggaggcg cgagggactg caggagcttc agggagtggc gcagctgctt catccccgtg
      541 gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt
      601 tctatgatga cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg
      661 cagtcggggc ggcgcggtcc caggtccact tcgcatatta aggtgacgcg tgtggcctcg
      721 aacaccgagc gaccctgcag cgacccgctt aaaagcttgg cattccggta ctgttggtaa
      781 agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
      841 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
      901 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
      961 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
     1021 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
     1081 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
     1141 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
     1201 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
     1261 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
     1321 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
     1381 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
     1441 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     1501 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     1561 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     1621 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     1681 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     1741 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     1801 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     1861 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     1921 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     1981 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     2041 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     2101 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     2161 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     2221 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     2281 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     2341 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     2401 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagcact agtaaagcaa
     2461 aaatgtgatc ttgcttgtaa atacaatttt gagaggttaa taaattacaa gtagtgctat
     2521 ttttgtattt aggttagcta tttagcttta cgttccagga tgcctagtgg cagccccaca
     2581 atatccagga agccctctct gcggtttttc agattaggta gtcgaaaaac ctaagaaatt
     2641 tacctgctac atttcaagat aatctagcca ccatgacttc gaaagtttat gatccagaac
     2701 aaaggaaacg gatgataact ggtccgcagt ggtgggccag atgtaaacaa atgaatgttc
     2761 ttgattcatt tattaattat tatgattcag aaaaacatgc agaaaatgct gttatttttt
     2821 tacatggtaa cgcggcctct tcttatttat ggcgacatgt tgtgccacat attgagccag
     2881 tagcgcggtg tattatacca gaccttattg gtatgggcaa atcaggcaaa tctggtaatg
     2941 gttcttatag gttacttgat cattacaaat atcttactgc atggtttgaa cttcttaatt
     3001 taccaaagaa gatcattttt gtcggccatg attggggtgc ttgtttggca tttcattata
     3061 gctatgagca tcaagataag atcaaagcaa tagttcacgc tgaaagtgta gtagatgtga
     3121 ttgaatcatg ggatgaatgg cctgatattg aagaagatat tgcgttgatc aaatctgaag
     3181 aaggagaaaa aatggttttg gagaataact tcttcgtgga aaccatgttg ccatcaaaaa
     3241 tcatgagaaa gttagaacca gaagaatttg cagcatatct tgaaccattc aaagagaaag
     3301 gtgaagttcg tcgtccaaca ttatcatggc ctcgtgaaat cccgttagta aaaggtggta
     3361 aacctgacgt tgtacaaatt gttaggaatt ataatgctta tctacgtgca agtgatgatt
     3421 taccaaaaat gtttattgaa tcggacccag gattcttttc caatgctatt gttgaaggtg
     3481 ccaagaagtt tcctaatact gaatttgtca aagtaaaagg tcttcatttt tcgcaagaag
     3541 atgcacctga tgaaatggga aaatatatca aatcgttcgt tgagcgagtt ctcaaaaatg
     3601 aacaataatt ctagactgta gaaaacatga ggatcaccca tgtctgctgg acgactgtag
     3661 aaaacatgag gatcacccat gtctgctgtc tagctgtaga aaacatgagg atcacccatg
     3721 tctgctggac gactgtagaa aacatgagga tcacccatgt ctgctgtcta gctgtagaaa
     3781 acatgaggat cacccatgtc tgctggacga ctgtagaaaa catgaggatc acccatgtct
     3841 gctgggccct agagcggccg cgattcgtca gtagggttgt aaaggttttt cttttcctga
     3901 gaaaacaacc ttttgttttc tcaggttttg ctttttggcc tttccctagc tttaaaaaaa
     3961 aaaaagcaaa agacgctggt ggctggcact cctggtttcc aggacggggt tcaagtccct
     4021 gcggtgtctt tgcttggatc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4081 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4141 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4261 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4381 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4441 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4561 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4741 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5041 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5341 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5821 cctggccttt tgctggcctt ttgctcacat ggctcgacag atcta
//
