LOCUS       sequence-327557-        3538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-421 and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3538)
  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 3538)
  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-mir421-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3538
                     /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    944..1025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1036..1053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1207..1795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1287..1306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1966..2757,2758..2826))
                     /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     2589..2608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2827..2931)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2964..3045
                     /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 cgcctatggc tctcgtctac cagactttaa
      601 attccttaag ggcaaggaca gtgccttact catctttgta ttcacagtgc ctaatccggt
      661 gcacattgta ggcctcatta aatgtttgtt gaatgaaaaa atgaatcatc aacagacatt
      721 aattgggcgc ctgctctgtg atctccatgg gctcagcttg tccccgccag ttgccaacaa
      781 cgtccaagct ctcttcagaa tgcttactcc tgaagcttgt cgacgatatc tccagaggat
      841 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
      901 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
      961 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1021 actgccccga gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2581 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2941 ccgaaaagtg ccacctgacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
     3001 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3061 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3121 ctctacaaat gtggtatggc tgattatgat cctctagact gcagggctgt cggccgcggc
     3181 ggcccccgtg gtgtggagtg cccctcagtg cggcgagaga ggtcccggcc tgagatacat
     3241 acttctttac attccatagc ttagcaggtc catatgggca tataaagaag tatgtttccc
     3301 aagcaggcag tctgcatgca gttccaagac cccgggacgc cgccccggta gaagctgacc
     3361 ccagatctgg gcccccgcgg catatgaccg gtgaattctc caggcgatct gacggttcac
     3421 taaacgagct ctgcttatat aggcctccca ccgtacacgc cacctcgaca tactcgagta
     3481 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
