LOCUS       sequence-327554-        3481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-181b-1 and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3481)
  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 3481)
  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-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      445..1033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     934..953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1187..1204
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1215..1296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1675..1878)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1704..1724)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1879..2182
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    2676..2757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2790..2894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2895..2963,2964..3481,1..274)
                     /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     complement(3113..3132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
       61 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      121 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      181 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      241 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      301 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
      361 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
      421 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      481 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      541 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      601 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      661 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      721 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      781 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      841 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      901 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      961 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1021 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1081 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1141 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1201 cgaggaagct cggggcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1261 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1321 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     1381 tgtggtatgg ctgattatga tcctctggag atatcgtcga caagcttctt tcaaagatgg
     1441 aatatctgtt gattgtgaca gcaaaaagcg gggccacagt tgcattcatt gttcagtgag
     1501 cttgtccaca cagttcaacc caccgacagc aatgaatgtt gattgtgacc ttttaaaaaa
     1561 taatctctgc acagggaaga gaaagtcctg gtgtgtccac ctttggtttg cggccgcgct
     1621 agcacgcgtc agctgactag aggatcccta gcggatctga cggttcacta aaccagctct
     1681 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     1741 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     1801 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     1861 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     1921 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     1981 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     2041 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     2101 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     2161 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     2221 aattgattac tattaataac tactcgagta tgtcgaggtg gcgtgtacgg tgggaggcct
     2281 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagaattcac cggtcatatg
     2341 ccgcgggggc ccagatctgg ggtcagcttc taccggggcg gcgtcccggg gtcttggaac
     2401 tgcatgcaga ctgcctgctt gggaaacata cttctttata tgcccatatg gacctgctaa
     2461 gctatggaat gtaaagaagt atgtatctca ggccgggacc tctctcgccg cactgagggg
     2521 cactccacac cacgggggcc gccgcggccg acagccctgc agtctagagg atcataatca
     2581 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2641 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2701 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgccga
     2761 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2821 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2881 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2941 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3001 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3061 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3121 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3181 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3241 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3301 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3361 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3421 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3481 t
//
