LOCUS       sequence-97816-e        4475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter to measure miR-155 mediated differential 
            repression. Contains PELO 3' UTR and mutated miR-155 sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4475)
  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 4475)
  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 mPELO
FEATURES             Location/Qualifiers
     source          1..4475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1354..1475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1365..1384)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1419..1438
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1608..1712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1713..1781,1782..2573)
                     /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(1931..1950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2744..3332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3233..3252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3407..4158
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4228..4360
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4343..4362
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4405..4423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4433..4475,1..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 cacactgcat gctttgaaac
      961 agttttcctt agaaaaagct tttgctatct tatcctgttt acattatttc tttattttaa
     1021 ttctgcttgg tgttcttgca ttgcatttaa tgatcccttt tctccccacc tccacacact
     1081 acattttttt tagatttaaa tagttttact attttaaatg attgccgtac aattagtaga
     1141 cttgaagaca agttttaaat atttttcttc aaaggcttgt taaaccaatc atgttaaaag
     1201 gaaattcttg gttttggttt gttgttgtta acgtaagtca tatttgattt agagggtaac
     1261 ttaaatcagt tatttttagc tttttagaac tttgatctgc tagggattgt caaaataatc
     1321 tccttgaggc gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     1381 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1441 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1501 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1561 gtggtaaaat cgataaggat ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1621 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1681 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1741 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1801 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1861 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1921 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1981 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2041 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2101 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2161 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2221 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2281 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2341 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2401 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2461 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2521 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2581 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2641 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2701 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2761 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2821 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2881 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2941 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3001 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3061 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3121 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3181 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3241 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3301 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3361 tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatctaaatg agtcttcgga
     3421 cctcgcgggg gccgcttaag cggtggttag ggtttgtctg acgcgggggg agggggaagg
     3481 aacgaaacac tctcattcgg aggcggctcg gggtttggtc ttggtggcca cgggcacgca
     3541 gaagagcgcc gcgatcctct taagcacccc cccgccctcc gtggaggcgg gggtttggtc
     3601 ggcgggtggt aactggcggg ccgctgactc gggcgggtcg cgcgccccag agtgtgacct
     3661 tttcggtctg ctcgcagacc cccgggcggc gccgccgcgg cggcgacggg ctcgctgggt
     3721 cctaggctcc atggggaccg tatacgtgga caggctctgg agcatccgca cgactgcggt
     3781 gatattaccg gagaccttct gcgggacgag ccgggtcacg cggctgacgc ggagcgtccg
     3841 ttgggcgaca aacaccagga cggggcacag gtacactatc ttgtcacccg gaggcgcgag
     3901 ggactgcagg agcttcaggg agtggcgcag ctgcttcatc cccgtggccc gttgctcgcg
     3961 tttgctggcg gtgtccccgg aagaaatata tttgcatgtc tttagttcta tgatgacaca
     4021 aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt cggggcggcg
     4081 cggtcccagg tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc
     4141 ctgcagcgac ccgcttaaaa gcttgattct tctgacacaa cagtctcgaa cttaagctgc
     4201 agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
     4261 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     4321 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat
     4381 tacagctctt aaggctagag tacttaatac gactcactat aggctagcca ccatgacttc
     4441 gaaagtttat gatccagaac aaaggaaacg gatga
//
