LOCUS       sequence-112558-        4530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter to measure miR-155 mediated differential 
            repression. Contains LMBRD2 3' UTR and mutated miR-155 sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4530)
  AUTHORS   Nam JW, Rissland OS, Koppstein D, Abreu-Goodger C, Jan CH, Agarwal 
            V, Yildirim MA, Rodriguez A, Bartel DP
  TITLE     Global analyses of the effect of different cellular contexts on 
            microRNA targeting.
  JOURNAL   Mol Cell. 2014 Mar 20;53(6):1031-43. doi: 
            10.1016/j.molcel.2014.02.013. Epub 2014 Mar 13.
  PUBMED    24631284
REFERENCE   2  (bases 1 to 4530)
  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: 
            "10.1016/j.molcel.2014.02.013"; journalName: "Mol Cell"; date: 
            "2014-03-20- 20"; volume: "53"; issue: "6"; pages: "1031-43"
COMMENT     Sequence Label: pIS1 mLMBRD2
FEATURES             Location/Qualifiers
     source          1..4530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1409..1530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1420..1439)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1474..1493
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1663..1767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1768..1836,1837..2628)
                     /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(1986..2005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2799..3387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3288..3307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3462..4213
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4283..4415
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4398..4417
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4460..4478
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4488..4530,1..893)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 taactggtcc gcagtggtgg gccagatgta aacaaatgaa tgttcttgat tcatttatta
       61 attattatga ttcagaaaaa catgcagaaa atgctgttat ttttttacat ggtaacgcgg
      121 cctcttctta tttatggcga catgttgtgc cacatattga gccagtagcg cggtgtatta
      181 taccagacct tattggtatg ggcaaatcag gcaaatctgg taatggttct tataggttac
      241 ttgatcatta caaatatctt actgcatggt ttgaacttct taatttacca aagaagatca
      301 tttttgtcgg ccatgattgg ggtgcttgtt tggcatttca ttatagctat gagcatcaag
      361 ataagatcaa agcaatagtt cacgctgaaa gtgtagtaga tgtgattgaa tcatgggatg
      421 aatggcctga tattgaagaa gatattgcgt tgatcaaatc tgaagaagga gaaaaaatgg
      481 ttttggagaa taacttcttc gtggaaacca tgttgccatc aaaaatcatg agaaagttag
      541 aaccagaaga atttgcagca tatcttgaac cattcaaaga gaaaggtgaa gttcgtcgtc
      601 caacattatc atggcctcgt gaaatcccgt tagtaaaagg tggtaaacct gacgttgtac
      661 aaattgttag gaattataat gcttatctac gtgcaagtga tgatttacca aaaatgttta
      721 ttgaatcgga cccaggattc ttttccaatg ctattgttga aggtgccaag aagtttccta
      781 atactgaatt tgtcaaagta aaaggtcttc atttttcgca agaagatgca cctgatgaaa
      841 tgggaaaata tatcaaatcg ttcgttgagc gagttctcaa aaatgaacaa taattctagg
      901 agctctatac cggtctcgat atcactacta gtgttctaga cttcaagtga tgcgtgtctg
      961 tgtatttagc atgtgtacta taatcaggtg atatagtatt ccttcagtct ttgtagtaac
     1021 tggatttttt tatgcttctg gtattgcttt ataaaagatt ttcatttcag aaagctatgt
     1081 tcacatttat catttaacat attgatttaa atgttgatta tgatattcaa ggataaatcc
     1141 tagacgtaaa tgaaaaatag ttttttttta agtagtcaaa gagtccaaca aaatccttca
     1201 gtatttaaag gtacctgctt atactttttg agaaaaaaac tggttattca aaggtaaaaa
     1261 gtgaatccat ttactgttat tactgatact taaatcgtgg tttatagaaa acaaaactat
     1321 attagaatgg aaatgtgtaa taacttgttg gttaaacatt cttactaaat agactgtttt
     1381 taaccgcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     1441 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1501 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1561 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     1621 aaaatcgata aggatccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1681 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1741 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1801 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1861 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1921 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1981 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2041 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2101 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2161 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2221 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2281 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2341 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2401 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2461 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2521 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2581 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2641 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2701 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2761 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2821 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2881 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2941 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3001 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3061 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3121 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3181 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3241 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3301 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3361 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3421 gccttttgct ggccttttgc tcacatggct cgacagatct aaatgagtct tcggacctcg
     3481 cgggggccgc ttaagcggtg gttagggttt gtctgacgcg gggggagggg gaaggaacga
     3541 aacactctca ttcggaggcg gctcggggtt tggtcttggt ggccacgggc acgcagaaga
     3601 gcgccgcgat cctcttaagc acccccccgc cctccgtgga ggcgggggtt tggtcggcgg
     3661 gtggtaactg gcgggccgct gactcgggcg ggtcgcgcgc cccagagtgt gaccttttcg
     3721 gtctgctcgc agacccccgg gcggcgccgc cgcggcggcg acgggctcgc tgggtcctag
     3781 gctccatggg gaccgtatac gtggacaggc tctggagcat ccgcacgact gcggtgatat
     3841 taccggagac cttctgcggg acgagccggg tcacgcggct gacgcggagc gtccgttggg
     3901 cgacaaacac caggacgggg cacaggtaca ctatcttgtc acccggaggc gcgagggact
     3961 gcaggagctt cagggagtgg cgcagctgct tcatccccgt ggcccgttgc tcgcgtttgc
     4021 tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc
     4081 cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc
     4141 ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca
     4201 gcgacccgct taaaagcttg attcttctga cacaacagtc tcgaacttaa gctgcagaag
     4261 ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca
     4321 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
     4381 tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag
     4441 ctcttaaggc tagagtactt aatacgactc actataggct agccaccatg acttcgaaag
     4501 tttatgatcc agaacaaagg aaacggatga
//
