LOCUS       sequence-112560-        4566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter to measure miR-155 mediated differential 
            repression. Contains EPAS1 3' UTR and mutated miR-155 sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4566)
  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 4566)
  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 mEPAS1
FEATURES             Location/Qualifiers
     source          1..4566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1445..1566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1456..1475)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1510..1529
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1699..1803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1804..1872,1873..2664)
                     /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(2022..2041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2835..3423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3324..3343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3498..4249
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4319..4451
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4434..4453
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4496..4514
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4524..4566,1..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 ctttgcaact ccctgggtaa
      961 gagggacgac acctctggtt tttcaatacc aattacatgg aacttttctg taatgggtac
     1021 aatgaagaag tttctaaaaa cacacacaaa gcacattggg ccaactattt agtaagcccg
     1081 gatagactta ttgccaaaaa caaaaaatag ctttcaaaag aaatttaagt tctatgagaa
     1141 attccttagt catggtgttg cgtaaatcat attttagctg cacgacgtaa ccccacacag
     1201 ggtggcagaa cttgaagggt tactgacgtg taaatgctgg tatttgattt cctgtgtgtg
     1261 ttgccctgac gtaaagggca ttttaccctt gcagttttac taaaacactg aaaaatattc
     1321 caagcttcat attaacccta cctgtcaacg taacgatttc atgaacgtta ttatattgtc
     1381 gaattcctac tgacaacatt ataactgtat gggagcttaa cgcggccgct tcgagcagac
     1441 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1501 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1561 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1621 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccaggtggc
     1681 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1741 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1801 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1861 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1921 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1981 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2041 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2101 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2161 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2221 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2281 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2341 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2401 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2461 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2521 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2581 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2641 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2701 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2761 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2821 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2881 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2941 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     3001 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3061 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3121 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3181 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3241 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3301 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3361 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3421 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3481 atggctcgac agatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta
     3541 gggtttgtct gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc
     3601 ggggtttggt cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc
     3661 ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact
     3721 cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg
     3781 cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg
     3841 acaggctctg gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga
     3901 gccgggtcac gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca
     3961 ggtacactat cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca
     4021 gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat
     4081 atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga
     4141 attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg
     4201 tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttgattc
     4261 ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     4321 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     4381 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     4441 tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata
     4501 cgactcacta taggctagcc accatgactt cgaaagttta tgatccagaa caaaggaaac
     4561 ggatga
//
