LOCUS       sequence-327555-        3765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-181b-1~125a and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3765)
  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 3765)
  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-mir181b-1~125-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3765
                     /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    1171..1252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1263..1280)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1434..2022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1514..1533)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2193..2984,2985..3053))
                     /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     2816..2835
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3054..3158)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3191..3272
                     /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 cgcaaaccaa aggtggacac accaggactt
      601 tctcttccct gtgcagagat tattttttaa aaggtcacaa tcaacattca ttgctgtcgg
      661 tgggttgaac tgtgtggaca agctcactga acaatgaatg caactgtggc cccgcttttt
      721 gctgtcacaa tcaacagata ttccatcttt gaaagcgttt ttggtctttc tgtctctggc
      781 tctcagaatg tctctgtgcc tatctccatc tctgaccccc accccagggt ctaccgggcc
      841 accgcacacc atgttgccag tctctaggtc cctgagaccc tttaacctgt gaggacatcc
      901 agggtcacag gtgaggttct tgggagcctg gcgtctggcc caaccacaca cctggggaat
      961 tgctggcctg acttctgacc cctgactcct catacccttc ctccagagca tgacatttga
     1021 ccaccaactg aaacctgaca agcttgtcga cgatatctcc agaggatcat aatcagccat
     1081 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1141 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1201 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gccccgagct
     1261 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1321 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1381 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1441 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1501 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1561 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1621 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1681 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1741 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1801 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1861 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1921 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1981 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2041 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2101 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2161 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2761 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     2821 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2881 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2941 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3001 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3061 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3121 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3181 cctgacgtcg gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3241 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3301 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3361 gtatggctga ttatgatcct ctagactgca gggctgtcgg ccgcggcggc ccccgtggtg
     3421 tggagtgccc ctcagtgcgg cgagagaggt cccggcctga gatacatact tctttacatt
     3481 ccatagctta gcaggtccat atgggcatat aaagaagtat gtttcccaag caggcagtct
     3541 gcatgcagtt ccaagacccc gggacgccgc cccggtagaa gctgacccca gatctgggcc
     3601 cccgcggcat atgaccggtg aattctccag gcgatctgac ggttcactaa acgagctctg
     3661 cttatatagg cctcccaccg tacacgccac ctcgacatac tcgagtagtt attaatagta
     3721 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
