LOCUS       sequence-97808-e        4585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter to measure miR-155 mediated differential 
            repression. Contains IL6R 3' UTR and mutated miR-155 sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4585)
  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 4585)
  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 mIL6R
FEATURES             Location/Qualifiers
     source          1..4585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1464..1585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1475..1494)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1529..1548
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1718..1822
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1823..1891,1892..2683)
                     /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(2041..2060)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2854..3442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3343..3362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3517..4268
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4338..4470
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4453..4472
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4515..4533
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4543..4585,1..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 gtgtctcaat gcacggggca
      961 tttctacctg tgtttctgca gcacccccac tgccttgagt ccccagcagt gctgttattt
     1021 gcctaacacc tgtagccatc tgccacgcag ccagacgtga aacgctgaga cagagaccat
     1081 ttaggttaaa tacgacagct tatcctgctg ggtggggaaa gtaaaaaata tgctggttca
     1141 aggcctaaag taaaatgatc aataatgttt gtaacgtaaa tgaaatattt tcaagaaatg
     1201 tgtccagggg tagcactggc tatgttgacg aggcctttgg taactcagag agctcttggc
     1261 cctgatgggg acttgccctt acgctttctt tatcaggctc tgagttcaca cggagcctct
     1321 ggcacttccc tgctgtcttg ggagaaagga aactggttgc cgcggcaggt tgtggaatct
     1381 gttgctggaa ccaggctgga agcccacctg gtagtgaaca gggcccagtg gggcaggctg
     1441 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1501 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1561 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1621 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     1681 cgataaggat ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1741 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1801 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1861 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1921 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1981 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2041 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2101 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2161 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2221 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2281 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2341 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2401 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2461 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2521 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2581 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2641 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2701 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2761 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2821 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2881 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2941 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3001 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3061 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3121 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3181 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3241 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3301 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3361 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3421 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3481 ttgctggcct tttgctcaca tggctcgaca gatctaaatg agtcttcgga cctcgcgggg
     3541 gccgcttaag cggtggttag ggtttgtctg acgcgggggg agggggaagg aacgaaacac
     3601 tctcattcgg aggcggctcg gggtttggtc ttggtggcca cgggcacgca gaagagcgcc
     3661 gcgatcctct taagcacccc cccgccctcc gtggaggcgg gggtttggtc ggcgggtggt
     3721 aactggcggg ccgctgactc gggcgggtcg cgcgccccag agtgtgacct tttcggtctg
     3781 ctcgcagacc cccgggcggc gccgccgcgg cggcgacggg ctcgctgggt cctaggctcc
     3841 atggggaccg tatacgtgga caggctctgg agcatccgca cgactgcggt gatattaccg
     3901 gagaccttct gcgggacgag ccgggtcacg cggctgacgc ggagcgtccg ttgggcgaca
     3961 aacaccagga cggggcacag gtacactatc ttgtcacccg gaggcgcgag ggactgcagg
     4021 agcttcaggg agtggcgcag ctgcttcatc cccgtggccc gttgctcgcg tttgctggcg
     4081 gtgtccccgg aagaaatata tttgcatgtc tttagttcta tgatgacaca aaccccgccc
     4141 agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt cggggcggcg cggtcccagg
     4201 tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac
     4261 ccgcttaaaa gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt
     4321 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
     4381 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
     4441 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt
     4501 aaggctagag tacttaatac gactcactat aggctagcca ccatgacttc gaaagtttat
     4561 gatccagaac aaaggaaacg gatga
//
