LOCUS       sequence-97806-e        4801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter to measure miR-155 mediated differential 
            repression. Contains HK1 3' UTR and mutated miR-155 sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4801)
  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 4801)
  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 mHK1
FEATURES             Location/Qualifiers
     source          1..4801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1680..1801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1691..1710)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1745..1764
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1934..2038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2039..2107,2108..2899)
                     /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(2257..2276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3070..3658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3559..3578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3733..4484
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4554..4686
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4669..4688
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4731..4749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4759..4801,1..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 gagtccggga tccccagcct
      961 actgcctctc cagcacttct ctcttcaagc ggcgaccccc taccctccca gcgagttgcg
     1021 ctgggagacg ctggcgccag ggcctgccgg cgcggggagg aaagcaaaat ccaactaatg
     1081 gtatatattg tagggtacag aatagagcgt gtgctgttga taatatctct cacccggatc
     1141 cctcctcact tgccctgcca ctttgcatgg tttgattttg acctggtccc ccacgtgtga
     1201 agtgtagtgg catccatttc taatgtatgc attcatccaa cagagttatt tattggctgg
     1261 agatggaaaa tcacaccacc tgacaggcct tctgggcctc caaagcccat ccttggggtt
     1321 ccccctccct gtgtgaaatg tattatcacc agcagacact gccgggcctc cctcccgggg
     1381 gcactgcctg aaggcgagtg tgggcataac gtaagctgct tcctcccctc ctggcaccca
     1441 ctgtggcctg gcatcgcatc gtggtgtgtc aatgccacaa aatcgtgtgt ccgtggaacc
     1501 agtcctagcc gcgtgtgaca gtcttgcatt ctgtttgtct cgtgggggga ggtggacagt
     1561 cctgcggaaa tgtgtcttgt ctccatttgg ataaaaggaa ccaaccaaca aacaatgcca
     1621 tcactggaat ttcccaccgc tttgtgagcc gtgtcggcgg ccgcttcgag cagacatgat
     1681 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1741 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1801 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1861 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccag gtggcacttt
     1921 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1981 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2041 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2101 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2161 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2221 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2281 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2341 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2401 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2461 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2521 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2581 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2641 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2701 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2761 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2821 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2881 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3061 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3121 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3181 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3541 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3601 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3661 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatggc
     3721 tcgacagatc taaatgagtc ttcggacctc gcgggggccg cttaagcggt ggttagggtt
     3781 tgtctgacgc ggggggaggg ggaaggaacg aaacactctc attcggaggc ggctcggggt
     3841 ttggtcttgg tggccacggg cacgcagaag agcgccgcga tcctcttaag cacccccccg
     3901 ccctccgtgg aggcgggggt ttggtcggcg ggtggtaact ggcgggccgc tgactcgggc
     3961 gggtcgcgcg ccccagagtg tgaccttttc ggtctgctcg cagacccccg ggcggcgccg
     4021 ccgcggcggc gacgggctcg ctgggtccta ggctccatgg ggaccgtata cgtggacagg
     4081 ctctggagca tccgcacgac tgcggtgata ttaccggaga ccttctgcgg gacgagccgg
     4141 gtcacgcggc tgacgcggag cgtccgttgg gcgacaaaca ccaggacggg gcacaggtac
     4201 actatcttgt cacccggagg cgcgagggac tgcaggagct tcagggagtg gcgcagctgc
     4261 ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg
     4321 catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg
     4381 aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat taaggtgacg
     4441 cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaaaagctt gattcttctg
     4501 acacaacagt ctcgaactta agctgcagaa gttggtcgtg aggcactggg caggtaagta
     4561 tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa
     4621 gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt gcctttctct
     4681 ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact taatacgact
     4741 cactataggc tagccaccat gacttcgaaa gtttatgatc cagaacaaag gaaacggatg
     4801 a
//
