LOCUS       sequence-354341-        6658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dual luminescence reporter to assess miRNA responsiveness in the 
            3'UTR of mouse Mov10 RNA helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6658)
  AUTHORS   Arora R, Bodak M, Penouty L, Hackman C, Ciaudo C
  TITLE     Sequestration of LINE-1 in cytosolic aggregates by MOV10 restricts 
            retrotransposition.
  JOURNAL   EMBO Rep. 2022 Jul 20:e54458. doi: 10.15252/embr.202154458.
  PUBMED    35856394
REFERENCE   2  (bases 1 to 6658)
  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"; journalName: "EMBO Rep. 
            2022 Jul 20:e54458. doi: 10.15252/embr.202154458."
COMMENT     Sequence Label: psiCHECK2-mMOV10-3'UTR-MRE16-mut1
FEATURES             Location/Qualifiers
     source          1..6658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2073..2121
                     /note="synthetic polyadenylation signal"
     promoter        2130..2881
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2911..2920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2917..4569
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3876..3884
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4613..4734
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4624..4643)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4678..4697
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4867..4971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4972..5040,5041..5832)
                     /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(5190..5209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6003..6591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6492..6511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgagcc cgggaattcg tttaaaccta gagcggccgc
     1681 gggaattcga tttaggcgat cgctcgaggc cacagccgcc cgccttctca caccagccaa
     1741 gccttattgc tgcctatccc tgacccagaa cccagctctc ctgtccctgc aagagatggg
     1801 aaggccggga agtttacagc ccagaccatt ccgccccttc ctctgctagg gaagacaacc
     1861 caagagtcta aaactcggag gaagggggag gaagaaagct tttttttttt tttttaaacc
     1921 ctctcctttg cagtgctggg gatcgaaccc agagcctcaa atatgctagg caagcactct
     1981 gcccctgagc tgctttaaga aaaacaaaat gctgttctat gcaaaaggcc gctggccgca
     2041 aaatcactag tgaattcgcg gccgctggcc gcaataaaat atctttattt tcattacatc
     2101 tgtgtgttgg ttttttgtgt gaggatctaa atgagtcttc ggacctcgcg ggggccgctt
     2161 aagcggtggt tagggtttgt ctgacgcggg gggaggggga aggaacgaaa cactctcatt
     2221 cggaggcggc tcggggtttg gtcttggtgg ccacgggcac gcagaagagc gccgcgatcc
     2281 tcttaagcac ccccccgccc tccgtggagg cgggggtttg gtcggcgggt ggtaactggc
     2341 gggccgctga ctcgggcggg tcgcgcgccc cagagtgtga ccttttcggt ctgctcgcag
     2401 acccccgggc ggcgccgccg cggcggcgac gggctcgctg ggtcctaggc tccatgggga
     2461 ccgtatacgt ggacaggctc tggagcatcc gcacgactgc ggtgatatta ccggagacct
     2521 tctgcgggac gagccgggtc acgcggctga cgcggagcgt ccgttgggcg acaaacacca
     2581 ggacggggca caggtacact atcttgtcac ccggaggcgc gagggactgc aggagcttca
     2641 gggagtggcg cagctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc
     2701 cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct
     2761 tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc aggtccactt
     2821 cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta
     2881 aaagcttggc attccggtac tgttggtaaa gccaccatgg ccgatgctaa gaacattaag
     2941 aagggccctg ctcccttcta ccctctggag gatggcaccg ctggcgagca gctgcacaag
     3001 gccatgaaga ggtatgccct ggtgcctggc accattgcct tcaccgatgc ccacattgag
     3061 gtggacatca cctatgccga gtacttcgag atgtctgtgc gcctggccga ggccatgaag
     3121 aggtacggcc tgaacaccaa ccaccgcatc gtggtgtgct ctgagaactc tctgcagttc
     3181 ttcatgccag tgctgggcgc cctgttcatc ggagtggccg tggcccctgc taacgacatt
     3241 tacaacgagc gcgagctgct gaacagcatg ggcatttctc agcctaccgt ggtgttcgtg
     3301 tctaagaagg gcctgcagaa gatcctgaac gtgcagaaga agctgcctat catccagaag
     3361 atcatcatca tggactctaa gaccgactac cagggcttcc agagcatgta cacattcgtg
     3421 acatctcatc tgcctcctgg cttcaacgag tacgacttcg tgccagagtc tttcgacagg
     3481 gacaaaacca ttgccctgat catgaacagc tctgggtcta ccggcctgcc taagggcgtg
     3541 gccctgcctc atcgcaccgc ctgtgtgcgc ttctctcacg cccgcgaccc tattttcggc
     3601 aaccagatca tccccgacac cgctattctg agcgtggtgc cattccacca cggcttcggc
     3661 atgttcacca ccctgggcta cctgatttgc ggctttcggg tggtgctgat gtaccgcttc
     3721 gaggaggagc tgttcctgcg cagcctgcaa gactacaaaa ttcagtctgc cctgctggtg
     3781 ccaaccctgt tcagcttctt cgctaagagc accctgatcg acaagtacga cctgtctaac
     3841 ctgcacgaga ttgcctctgg cggcgcccca ctgtctaagg aggtgggcga agccgtggcc
     3901 aagcgctttc atctgccagg catccgccag ggctacggcc tgaccgagac aaccagcgcc
     3961 attctgatta ccccagaggg cgacgacaag cctggcgccg tgggcaaggt ggtgccattc
     4021 ttcgaggcca aggtggtgga cctggacacc ggcaagaccc tgggagtgaa ccagcgcggc
     4081 gagctgtgtg tgcgcggccc tatgattatg tccggctacg tgaataaccc tgaggccaca
     4141 aacgccctga tcgacaagga cggctggctg cactctggcg acattgccta ctgggacgag
     4201 gacgagcact tcttcatcgt ggaccgcctg aagtctctga tcaagtacaa gggctaccag
     4261 gtggccccag ccgagctgga gtctatcctg ctgcagcacc ctaacatttt cgacgccgga
     4321 gtggccggcc tgcccgacga cgatgccggc gagctgcctg ccgccgtcgt cgtgctggaa
     4381 cacggcaaga ccatgaccga gaaggagatc gtggactatg tggccagcca ggtgacaacc
     4441 gccaagaagc tgcgcggcgg agtggtgttc gtggacgagg tgcccaaggg cctgaccggc
     4501 aagctggacg cccgcaagat ccgcgagatc ctgatcaagg ctaagaaagg cggcaagatc
     4561 gccgtgtaat aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     4621 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4681 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4741 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     4801 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc caggtggcac ttttcgggga
     4861 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4921 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4981 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5041 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5101 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5161 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5221 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5281 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5341 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5401 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5461 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5521 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5581 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5641 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5701 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5761 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5821 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5881 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5941 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat ggctcgac
//
