LOCUS       sequence-112562-        4590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter to measure miR-155 mediated differential 
            repression. Contains INSIG2 3' UTR and mutated miR-155 sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4590)
  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 4590)
  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 mINSIG2
FEATURES             Location/Qualifiers
     source          1..4590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1469..1590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1480..1499)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1534..1553
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1723..1827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1828..1896,1897..2688)
                     /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(2046..2065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2859..3447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3348..3367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3522..4273
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4343..4475
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4458..4477
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4520..4538
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4548..4590,1..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 gtattttctc tttagcttac
      961 tgttgtgcct ttttattttt ctaatcacag cagtatgagt tatgagtgcc ctaatttgtg
     1021 gttagtttct aatttaatgt tgtttcatag agtttggagt gttttgatac agggtgaaaa
     1081 tgaacttctg gtttcaaacc tgcgttactg gagacagccc aaagagtaat tttctgtttt
     1141 gacaggtttt actggaagta tatgtgatga gcagaagagg ttatcaacgt caaattgttt
     1201 tggttctaaa tttggaacag tatatataat taaaagtaag gaacattaga ggatttaatt
     1261 agaataaata catgttttgg aaatacagtg acctcttgca gtgtcacaaa agtgcaaagt
     1321 gatattagct gtcatctgca atacagaatc tcattgcttt tgcacatgga gcatatagga
     1381 aactccaaac agatcacaat gaggtttcta aatctgttgg gttctgtctt ctattgggtt
     1441 ctgtggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     1501 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1561 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1621 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     1681 aaaatcgata aggatccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1801 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1861 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1921 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1981 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2041 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2101 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2161 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2221 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2281 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2341 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2401 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2461 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2521 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2581 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2641 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2701 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2761 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2821 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2881 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2941 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3001 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3061 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3121 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3181 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3241 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3301 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3361 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3421 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3481 gccttttgct ggccttttgc tcacatggct cgacagatct aaatgagtct tcggacctcg
     3541 cgggggccgc ttaagcggtg gttagggttt gtctgacgcg gggggagggg gaaggaacga
     3601 aacactctca ttcggaggcg gctcggggtt tggtcttggt ggccacgggc acgcagaaga
     3661 gcgccgcgat cctcttaagc acccccccgc cctccgtgga ggcgggggtt tggtcggcgg
     3721 gtggtaactg gcgggccgct gactcgggcg ggtcgcgcgc cccagagtgt gaccttttcg
     3781 gtctgctcgc agacccccgg gcggcgccgc cgcggcggcg acgggctcgc tgggtcctag
     3841 gctccatggg gaccgtatac gtggacaggc tctggagcat ccgcacgact gcggtgatat
     3901 taccggagac cttctgcggg acgagccggg tcacgcggct gacgcggagc gtccgttggg
     3961 cgacaaacac caggacgggg cacaggtaca ctatcttgtc acccggaggc gcgagggact
     4021 gcaggagctt cagggagtgg cgcagctgct tcatccccgt ggcccgttgc tcgcgtttgc
     4081 tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc
     4141 cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc
     4201 ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca
     4261 gcgacccgct taaaagcttg attcttctga cacaacagtc tcgaacttaa gctgcagaag
     4321 ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca
     4381 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
     4441 tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag
     4501 ctcttaaggc tagagtactt aatacgactc actataggct agccaccatg acttcgaaag
     4561 tttatgatcc agaacaaagg aaacggatga
//
