LOCUS       sequence-327556-        3550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-374b and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3550)
  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 3550)
  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-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3550
                     /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    956..1037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1048..1065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1219..1807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1299..1318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1978..2769,2770..2838))
                     /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     2601..2620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2839..2943)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2976..3057
                     /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 ggtgaagctt gtcgacgata
      841 tctccagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
      901 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
      961 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1021 agcatttttt tcactgcccc gagcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1621 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2581 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2821 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2941 gcgcacattt ccccgaaaag tgccacctga cgtcggcagt gaaaaaaatg ctttatttgt
     3001 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3061 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3121 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga ctgcagggct
     3181 gtcggccgcg gcggcccccg tggtgtggag tgcccctcag tgcggcgaga gaggtcccgg
     3241 cctgagatac atacttcttt acattccata gcttagcagg tccatatggg catataaaga
     3301 agtatgtttc ccaagcaggc agtctgcatg cagttccaag accccgggac gccgccccgg
     3361 tagaagctga ccccagatct gggcccccgc ggcatatgac cggtgaattc tccaggcgat
     3421 ctgacggttc actaaacgag ctctgcttat ataggcctcc caccgtacac gccacctcga
     3481 catactcgag tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     3541 tggagttccg
//
