LOCUS       sequence-203168-        8088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the 3'UTR of relevant gene for validation of miRNAs 
            targeting the 3'UTR in a dual luciferase assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8088)
  AUTHORS   Ramalingam D, Ziegelbauer JM
  TITLE     Viral microRNAs Target a Gene Network, Inhibit STAT Activation, and 
            Suppress Interferon Responses.
  JOURNAL   Sci Rep. 2017 Jan 19;7:40813. doi: 10.1038/srep40813.
  PUBMED    28102325
REFERENCE   2  (bases 1 to 8088)
  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"; journalName: "Sci Rep."; 
            date: "2017-01-19"; volume: "7:40813. doi"; pages: " 
            10.1038/srep40813"
COMMENT     Sequence Label: pD765 HMGCR 3'UTR
FEATURES             Location/Qualifiers
     source          1..8088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        85..442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        85..276
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(108..128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      293..428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          512..644
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     627..646
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        689..707
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      711..720
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             717..1652
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     protein_bind    1690..1714
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3474..3498)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3526..3574
                     /note="synthetic polyadenylation signal"
     promoter        3583..4334
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      4364..4373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4370..6022
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             5329..5337
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    6066..6187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6077..6096)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6131..6150
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6320..6424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6425..6493,6494..7285)
                     /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(6643..6662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7456..8044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7945..7964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct cacatggctc gacagatctg cgcagcacca tggcctgaaa taacctctga
       61 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
      121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      241 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      361 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
      421 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
      481 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      541 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      601 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
      661 caattacagc tcttaaggct agagtactta atacgactca ctataggcta gccaccatgg
      721 cttccaaggt gtacgacccc gagcaacgca aacgcatgat cactgggcct cagtggtggg
      781 ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa ctactatgat tccgagaagc
      841 acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc ctccagctac ctgtggaggc
      901 acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat ccctgatctg atcggaatgg
      961 gtaagtccgg caagagcggg aatggctcat atcgcctcct ggatcactac aagtacctca
     1021 ccgcttggtt cgagctgctg aaccttccaa agaaaatcat ctttgtgggc cacgactggg
     1081 gggcttgtct ggcctttcac tactcctacg agcaccaaga caagatcaag gccatcgtcc
     1141 atgctgagag tgtcgtggac gtgatcgagt cctgggacga gtggcctgac atcgaggagg
     1201 atatcgccct gatcaagagc gaagagggcg agaaaatggt gcttgagaat aacttcttcg
     1261 tcgagaccat gctcccaagc aagatcatgc ggaaactgga gcctgaggag ttcgctgcct
     1321 acctggagcc attcaaggag aagggcgagg ttagacggcc taccctctcc tggcctcgcg
     1381 agatccctct cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc aactacaacg
     1441 cctaccttcg ggccagcgac gatctgccta agatgttcat cgagtccgac cctgggttct
     1501 tttccaacgc tattgtcgag ggagctaaga agttccctaa caccgagttc gtgaaggtga
     1561 agggcctcca cttcagccag gaggacgctc cagatgaaat gggtaagtac atcaagagct
     1621 tcgtggagcg cgtgctgaag aacgagcagt aattctaggc gatcgctcga gcccgggaat
     1681 tcgtttatca caagtttgta caaaaaagtt ggcatagccc gacagttctg aactggaaca
     1741 tgggcattgg gttctaaagg actaacataa aatctgtgaa ttaaaaaagc tcaatgcatt
     1801 gtcttgtgga ggatgaatag atgtgatcac tgagacagcc acttggtttt tggctctttc
     1861 agagaggtct caggttcttt ccatgcagac tcctcagatc tgaacacagt ttagtgcttt
     1921 acatgctgtg ctctttgaag agatttcaac aagaatattg tatgttaaag catcagagat
     1981 ggtaatctac agctcacctc tgaaggcaaa tataagctgg gaaaaaagtt ttgatgaaat
     2041 tcttgaagtt catggtgatc agtgcaattg accttctccc tcactcctgc cagttgaaaa
     2101 tggattttta aattatactg tagctgatga aactcctgat tttgtagtta atttattaag
     2161 tctgggatgt agaacttcaa gaagtaagag ctaagttcta agttcatgtt tgtaaattaa
     2221 tacttcattt ggtgctggtc tattttgatt ttggggggta atcagcatta ttcttcagaa
     2281 ggggacctgt tttcttcaag ggaagaaaca ctcttattcc caaactacag aataatgtgt
     2341 taaacatgct aaatagttct atcaggaaaa caaatcactg tatttatctc cgcaggctat
     2401 ttgttcagag aggccttttg tttaaatata aatgtttaaa tataaatgtt tgtctggatt
     2461 ggctataaca tgtctttcag cattaggctt ttaagaaaca cagggttttg tattctttac
     2521 taaagatatc agagctctta atgttgctta gatgagggtg actgtcaagt acaagcaaga
     2581 ctgggacctt agaaatcatt gtagaaacac agttttgaaa gaaaaatacc atgtctctaa
     2641 gccaacttta attgcttaaa agacattttt atttagttga aaaatctagt tttttttgta
     2701 aactgtatca aatctgtata tgttgtaata aaacttatgc tagtttattg gaagtgttca
     2761 agaaataaaa atcaacttgt gtactgataa aatactctag cctgggccag agaagataat
     2821 gttctttaat gttgtccagg aaaccctggc ttgcttgccg agcctaatga aagggaaagt
     2881 cagctttcag agccagtgaa ggagccacgt gaatggccct agaactgtgc ctagttcctg
     2941 tggccaggag gttggtgact gaaacattca cacagggctc tttgatggac ccacgaacgc
     3001 tcttagcttt ctcagggggt cagcagagtt attgaatctt aatttttttt aatgtacaag
     3061 ttttgtataa ataataaaga actccttatt ttgtattaca tctaatgctt caagtgttgc
     3121 tcttggaaag ctgatgatgt ctcttgtaga agatggactc tgaaaaacat tccaggaaac
     3181 catggcagca tggagagcct cttagtgatt gtgtctgcat tgttattgtg gaagatttac
     3241 cttttctgtt gtacgtaaag cttaaattgc ttttgttgtg actttttagc cagtgacttt
     3301 ttctgagctt ttcatggaag tggcagtgaa aaatatgttg agtgttcatt ttagtgactg
     3361 taattaatat cttgctggat taatgttttg tacaattact aaattgtata cattttgtta
     3421 tagaatactt ttttctagtt tcagtaaata atgaaaagga agttaatacc aataatcaac
     3481 tttcttgtac aaagtggtga taaacctaga gcggccgctg gccgcaataa aatatcttta
     3541 ttttcattac atctgtgtgt tggttttttg tgtgaggatc taaatgagtc ttcggacctc
     3601 gcgggggccg cttaagcggt ggttagggtt tgtctgacgc ggggggaggg ggaaggaacg
     3661 aaacactctc attcggaggc ggctcggggt ttggtcttgg tggccacggg cacgcagaag
     3721 agcgccgcga tcctcttaag cacccccccg ccctccgtgg aggcgggggt ttggtcggcg
     3781 ggtggtaact ggcgggccgc tgactcgggc gggtcgcgcg ccccagagtg tgaccttttc
     3841 ggtctgctcg cagacccccg ggcggcgccg ccgcggcggc gacgggctcg ctgggtccta
     3901 ggctccatgg ggaccgtata cgtggacagg ctctggagca tccgcacgac tgcggtgata
     3961 ttaccggaga ccttctgcgg gacgagccgg gtcacgcggc tgacgcggag cgtccgttgg
     4021 gcgacaaaca ccaggacggg gcacaggtac actatcttgt cacccggagg cgcgagggac
     4081 tgcaggagct tcagggagtg gcgcagctgc ttcatccccg tggcccgttg ctcgcgtttg
     4141 ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc
     4201 ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt
     4261 cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc
     4321 agcgacccgc ttaaaagctt ggcattccgg tactgttggt aaagccacca tggccgatgc
     4381 taagaacatt aagaagggcc ctgctccctt ctaccctctg gaggatggca ccgctggcga
     4441 gcagctgcac aaggccatga agaggtatgc cctggtgcct ggcaccattg ccttcaccga
     4501 tgcccacatt gaggtggaca tcacctatgc cgagtacttc gagatgtctg tgcgcctggc
     4561 cgaggccatg aagaggtacg gcctgaacac caaccaccgc atcgtggtgt gctctgagaa
     4621 ctctctgcag ttcttcatgc cagtgctggg cgccctgttc atcggagtgg ccgtggcccc
     4681 tgctaacgac atttacaacg agcgcgagct gctgaacagc atgggcattt ctcagcctac
     4741 cgtggtgttc gtgtctaaga agggcctgca gaagatcctg aacgtgcaga agaagctgcc
     4801 tatcatccag aagatcatca tcatggactc taagaccgac taccagggct tccagagcat
     4861 gtacacattc gtgacatctc atctgcctcc tggcttcaac gagtacgact tcgtgccaga
     4921 gtctttcgac agggacaaaa ccattgccct gatcatgaac agctctgggt ctaccggcct
     4981 gcctaagggc gtggccctgc ctcatcgcac cgcctgtgtg cgcttctctc acgcccgcga
     5041 ccctattttc ggcaaccaga tcatccccga caccgctatt ctgagcgtgg tgccattcca
     5101 ccacggcttc ggcatgttca ccaccctggg ctacctgatt tgcggctttc gggtggtgct
     5161 gatgtaccgc ttcgaggagg agctgttcct gcgcagcctg caagactaca aaattcagtc
     5221 tgccctgctg gtgccaaccc tgttcagctt cttcgctaag agcaccctga tcgacaagta
     5281 cgacctgtct aacctgcacg agattgcctc tggcggcgcc ccactgtcta aggaggtggg
     5341 cgaagccgtg gccaagcgct ttcatctgcc aggcatccgc cagggctacg gcctgaccga
     5401 gacaaccagc gccattctga ttaccccaga gggcgacgac aagcctggcg ccgtgggcaa
     5461 ggtggtgcca ttcttcgagg ccaaggtggt ggacctggac accggcaaga ccctgggagt
     5521 gaaccagcgc ggcgagctgt gtgtgcgcgg ccctatgatt atgtccggct acgtgaataa
     5581 ccctgaggcc acaaacgccc tgatcgacaa ggacggctgg ctgcactctg gcgacattgc
     5641 ctactgggac gaggacgagc acttcttcat cgtggaccgc ctgaagtctc tgatcaagta
     5701 caagggctac caggtggccc cagccgagct ggagtctatc ctgctgcagc accctaacat
     5761 tttcgacgcc ggagtggccg gcctgcccga cgacgatgcc ggcgagctgc ctgccgccgt
     5821 cgtcgtgctg gaacacggca agaccatgac cgagaaggag atcgtggact atgtggccag
     5881 ccaggtgaca accgccaaga agctgcgcgg cggagtggtg ttcgtggacg aggtgcccaa
     5941 gggcctgacc ggcaagctgg acgcccgcaa gatccgcgag atcctgatca aggctaagaa
     6001 aggcggcaag atcgccgtgt aataattcta gagtcggggc ggccggccgc ttcgagcaga
     6061 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     6121 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     6181 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     6241 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccaggtgg
     6301 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6361 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6421 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6481 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6541 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6601 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6661 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6721 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6781 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6841 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6901 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6961 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7021 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7081 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7141 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7201 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7261 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7321 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7381 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7441 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7501 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7561 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7621 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7681 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7741 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7801 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7861 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7921 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7981 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8041 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctg
//
