LOCUS       sequence-203166-        7023 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 7023)
  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 7023)
  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 FDFT1 3'UTR
FEATURES             Location/Qualifiers
     source          1..7023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        61..252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          488..620
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     603..622
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        665..683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             693..1628
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     protein_bind    1666..1690
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2385..2409)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2437..2485
                     /note="synthetic polyadenylation signal"
     promoter        2494..3245
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3275..3284
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3281..4933
                     /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             4240..4248
                     /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    4977..5098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4988..5007)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5042..5061
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5231..5335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5336..5404,5405..6196)
                     /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(5554..5573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6367..6955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6856..6875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
       61 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac
      481 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
      541 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
      601 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
      661 tacttaatac gactcactat aggctagcca ccatggcttc caaggtgtac gaccccgagc
      721 aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc
      781 tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc gtgatttttc
      841 tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac atcgagcccg
      901 tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag agcgggaatg
      961 gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag ctgctgaacc
     1021 ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc tttcactact
     1081 cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga
     1141 tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc aagagcgaag
     1201 agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc ccaagcaaga
     1261 tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc aaggagaagg
     1321 gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt aagggaggca
     1381 agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc agcgacgatc
     1441 tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt gtcgagggag
     1501 ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc agccaggagg
     1561 acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg
     1621 agcagtaatt ctaggcgatc gctcgagccc gggaattcgt ttatcacaag tttgtacaaa
     1681 aaagttggct cccaaatttg tccatagctg aagtccacca taaagtggat ttactttttt
     1741 tctttaagga tggatgttgt gttctcttta tttttttcct actactttaa tccctaaaag
     1801 aacgctgtgt ggctgggacc tttaggaaag tgaaatgcag gtgagaagaa cctaaacatg
     1861 aaaggaaagg gtgcctcatc ccagcaacct gtccttgtgg gtgatgatca ctgtgctgct
     1921 tgcggctcat ggcagagcat tcagtgccac ggtttaggtg aagtcgctgc atatgtgact
     1981 gtcatgagat cctacttagt atgatcctgg ctagaatgat aattaaaagt atttaatttg
     2041 aagcaccatt tgaatgttcg taatagtaga aaatgatgtg aattttcttt ctgttcggct
     2101 cctatttttc tcatcatttt gttttcttta attgggttga atggagtaga tagaaatatt
     2161 tatggtttag gtaacagtta gatgtttcct aagaatgcaa actgcctttt ccacacaaag
     2221 gctgggaata aaattctggg tattctcgta ttctcattta aaggagttta gctttcagag
     2281 agaaacagca ggattgcttt tgacctttta gaagattggt ctccagtaaa ggtggacatt
     2341 tttgagattt ttataataaa gaatttaatt gctctgcatg tcaatatcaa ctttcttgta
     2401 caaagtggtg ataaacctag agcggccgct ggccgcaata aaatatcttt attttcatta
     2461 catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt cttcggacct cgcgggggcc
     2521 gcttaagcgg tggttagggt ttgtctgacg cggggggagg gggaaggaac gaaacactct
     2581 cattcggagg cggctcgggg tttggtcttg gtggccacgg gcacgcagaa gagcgccgcg
     2641 atcctcttaa gcaccccccc gccctccgtg gaggcggggg tttggtcggc gggtggtaac
     2701 tggcgggccg ctgactcggg cgggtcgcgc gccccagagt gtgacctttt cggtctgctc
     2761 gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc gctgggtcct aggctccatg
     2821 gggaccgtat acgtggacag gctctggagc atccgcacga ctgcggtgat attaccggag
     2881 accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga gcgtccgttg ggcgacaaac
     2941 accaggacgg ggcacaggta cactatcttg tcacccggag gcgcgaggga ctgcaggagc
     3001 ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg
     3061 tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc
     3121 gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc
     3181 acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg
     3241 cttaaaagct tggcattccg gtactgttgg taaagccacc atggccgatg ctaagaacat
     3301 taagaagggc cctgctccct tctaccctct ggaggatggc accgctggcg agcagctgca
     3361 caaggccatg aagaggtatg ccctggtgcc tggcaccatt gccttcaccg atgcccacat
     3421 tgaggtggac atcacctatg ccgagtactt cgagatgtct gtgcgcctgg ccgaggccat
     3481 gaagaggtac ggcctgaaca ccaaccaccg catcgtggtg tgctctgaga actctctgca
     3541 gttcttcatg ccagtgctgg gcgccctgtt catcggagtg gccgtggccc ctgctaacga
     3601 catttacaac gagcgcgagc tgctgaacag catgggcatt tctcagccta ccgtggtgtt
     3661 cgtgtctaag aagggcctgc agaagatcct gaacgtgcag aagaagctgc ctatcatcca
     3721 gaagatcatc atcatggact ctaagaccga ctaccagggc ttccagagca tgtacacatt
     3781 cgtgacatct catctgcctc ctggcttcaa cgagtacgac ttcgtgccag agtctttcga
     3841 cagggacaaa accattgccc tgatcatgaa cagctctggg tctaccggcc tgcctaaggg
     3901 cgtggccctg cctcatcgca ccgcctgtgt gcgcttctct cacgcccgcg accctatttt
     3961 cggcaaccag atcatccccg acaccgctat tctgagcgtg gtgccattcc accacggctt
     4021 cggcatgttc accaccctgg gctacctgat ttgcggcttt cgggtggtgc tgatgtaccg
     4081 cttcgaggag gagctgttcc tgcgcagcct gcaagactac aaaattcagt ctgccctgct
     4141 ggtgccaacc ctgttcagct tcttcgctaa gagcaccctg atcgacaagt acgacctgtc
     4201 taacctgcac gagattgcct ctggcggcgc cccactgtct aaggaggtgg gcgaagccgt
     4261 ggccaagcgc tttcatctgc caggcatccg ccagggctac ggcctgaccg agacaaccag
     4321 cgccattctg attaccccag agggcgacga caagcctggc gccgtgggca aggtggtgcc
     4381 attcttcgag gccaaggtgg tggacctgga caccggcaag accctgggag tgaaccagcg
     4441 cggcgagctg tgtgtgcgcg gccctatgat tatgtccggc tacgtgaata accctgaggc
     4501 cacaaacgcc ctgatcgaca aggacggctg gctgcactct ggcgacattg cctactggga
     4561 cgaggacgag cacttcttca tcgtggaccg cctgaagtct ctgatcaagt acaagggcta
     4621 ccaggtggcc ccagccgagc tggagtctat cctgctgcag caccctaaca ttttcgacgc
     4681 cggagtggcc ggcctgcccg acgacgatgc cggcgagctg cctgccgccg tcgtcgtgct
     4741 ggaacacggc aagaccatga ccgagaagga gatcgtggac tatgtggcca gccaggtgac
     4801 aaccgccaag aagctgcgcg gcggagtggt gttcgtggac gaggtgccca agggcctgac
     4861 cggcaagctg gacgcccgca agatccgcga gatcctgatc aaggctaaga aaggcggcaa
     4921 gatcgccgtg taataattct agagtcgggg cggccggccg cttcgagcag acatgataag
     4981 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     5041 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     5101 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     5161 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccaggtg gcacttttcg
     5221 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5281 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5341 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5401 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5461 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5521 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5581 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5641 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5701 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5761 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5821 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5881 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5941 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6001 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6061 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6121 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6181 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6241 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6301 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6361 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6421 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6481 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6541 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6601 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6661 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6721 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6781 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6841 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6901 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6961 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     7021 aca
//
