LOCUS       sequence-328512-        3479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-144 and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3479)
  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 3479)
  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-mir144-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        160..463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        464..667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     618..638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1044..1125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1136..1153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1307..1895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1387..1406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2066..2857,2858..2926))
                     /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     2689..2708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2927..3031)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3064..3145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gcatatgacc ggtgaattct ccaggcgatc tgacggttca ctaaacgagc tctgcttata
       61 taggcctccc accgtacacg ccacctcgac atactcgagt agttattaat agtaatcaat
      121 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      421 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      481 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      541 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      601 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      661 cagagctggt ttagtgaacc gtcagatccg ctagggatcc tctagtcagc tgacgcgtgc
      721 tagcgcggcc gcgttctgcc cccagcccct cacagtgctt ttcaagccat gcttcctgtg
      781 cccccagtgg ggccctggct gggatatcat catatactgt aagtttgcga tgagacacta
      841 cagtatagat gatgtactag tccgggcacc cccagctctg gagcctgaca aggaggacag
      901 gagagatgct gcaagcttgt cgacgatatc tccagaggat cataatcagc cataccacat
      961 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1021 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1081 gcaatagcat cacaaatttc acaaataaag catttttttc actgccccga gcttcctcgc
     1141 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1201 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1261 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1321 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1381 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1441 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1501 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1741 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     2701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2941 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3001 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3061 tcggcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3121 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3181 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     3241 tgattatgat cctctagact gcagggctgt cggccgcggc ggcccccgtg gtgtggagtg
     3301 cccctcagtg cggcgagaga ggtcccggcc tgagatacat acttctttac attccatagc
     3361 ttagcaggtc catatgggca tataaagaag tatgtttccc aagcaggcag tctgcatgca
     3421 gttccaagac cccgggacgc cgccccggta gaagctgacc ccagatctgg gcccccgcg
//
