LOCUS       sequence-327558-        3701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-374b~421 and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3701)
  AUTHORS   Fang W, Bartel DP
  TITLE     MicroRNA Clustering Assists Processing of Suboptimal MicroRNA 
            Hairpins through the Action of the ERH Protein.
  JOURNAL   Mol Cell. 2020 Apr 16;78(2):289-302.e6. doi: 
            10.1016/j.molcel.2020.01.026.
  PUBMED    32302541
REFERENCE   2  (bases 1 to 3701)
  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.2020.01"; journalName: "Mol Cell"; date: 
            "2020-04-16- 16"; volume: "78"; issue: "2"; pages: "289-302.e6"
COMMENT     Sequence Label: pBI-CMV1-mir374b~421-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1107..1188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1199..1216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1370..1958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1450..1469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2129..2920,2921..2989))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2752..2771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2990..3094)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3127..3208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagggatc
      541 ctctagtcag ctgacgcgtg ctagcgcggc cgcagcacgt gacaaaaaca acagcagacc
      601 ctggtgcctg ggaggacttc atggatccag cagcaacctg gagtggtgct cctctgaaga
      661 aatcctactc ggatggatat aatacaacct gctaagtgtc ctagcactta gcaggttgta
      721 ttatcattgt ccgtgtctat ggctctcgtc taccagactt taaattcctt aagggcaagg
      781 acagtgcctt actcatcttt gtattcacag tgcctaatcc ggtgcacatt gtaggcctca
      841 ttaaatgttt gttgaatgaa aaaatgaatc atcaacagac attaattggg cgcctgctct
      901 gtgatctcca tgggctcagc ttgtccccgc cagttgccaa caacgtccaa gctctcttca
      961 gaatgcttac tcctgaagct tgtcgacgat atctccagag gatcataatc agccatacca
     1021 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1081 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgccc cgagcttcct
     1201 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1261 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1561 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2041 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2101 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2161 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2221 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2281 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2341 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2401 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2461 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2521 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2581 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2641 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2701 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     2761 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2821 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2881 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2941 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3001 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3061 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3121 acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3181 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3241 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     3301 ggctgattat gatcctctag actgcagggc tgtcggccgc ggcggccccc gtggtgtgga
     3361 gtgcccctca gtgcggcgag agaggtcccg gcctgagata catacttctt tacattccat
     3421 agcttagcag gtccatatgg gcatataaag aagtatgttt cccaagcagg cagtctgcat
     3481 gcagttccaa gaccccggga cgccgccccg gtagaagctg accccagatc tgggcccccg
     3541 cggcatatga ccggtgaatt ctccaggcga tctgacggtt cactaaacga gctctgctta
     3601 tataggcctc ccaccgtaca cgccacctcg acatactcga gtagttatta atagtaatca
     3661 attacggggt cattagttca tagcccatat atggagttcc g
//
