LOCUS       sequence-328912-        3656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-451~144 and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3656)
  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 3656)
  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-mir451~144-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        140..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     598..618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1201..1282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1293..1310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1464..2052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1544..1563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2223..3014,3015..3083))
                     /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     2846..2865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3084..3188)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3221..3302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggcctccc accgtacacg
       61 ccacctcgac atactcgagt agttattaat agtaatcaat tacggggtca ttagttcata
      121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      661 gtcagatccg ctagggatcc tctagtcagc tgacgcgtgc tagcgcggcc gcaagaagct
      721 ctctgctcag cctgtcacaa cctactgact gccagggcac ttgggaatgg caaggaaacc
      781 gttaccatta ctgagtttag taatggtaat ggttctcttg ctatacccag aaaacgtgcc
      841 aggaagagaa ctcaggaccc tgaagcagac tactggaagg gagactccag ttctgccccc
      901 agcccctcac agtgcttttc aagccatgct tcctgtgccc ccagtggggc cctggctggg
      961 atatcatcat atactgtaag tttgcgatga gacactacag tatagatgat gtactagtcc
     1021 gggcaccccc agctctggag cctgacaagg aggacaggag agatgctgca agcttgtcga
     1081 cgatatctcc agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1141 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1201 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1261 aataaagcat ttttttcact gccccgagct tcctcgctca ctgactcgct gcgctcggtc
     1321 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1381 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1441 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1501 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1561 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1621 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1681 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1741 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1801 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1861 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1921 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1981 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2041 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2101 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2161 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2221 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2281 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2341 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2401 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2461 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2521 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2581 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2641 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2701 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2761 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2821 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     2881 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2941 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3001 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3061 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3121 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3181 gttccgcgca catttccccg aaaagtgcca cctgacgtcg gcagtgaaaa aaatgcttta
     3241 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3301 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3361 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcct ctagactgca
     3421 gggctgtcgg ccgcggcggc ccccgtggtg tggagtgccc ctcagtgcgg cgagagaggt
     3481 cccggcctga gatacatact tctttacatt ccatagctta gcaggtccat atgggcatat
     3541 aaagaagtat gtttcccaag caggcagtct gcatgcagtt ccaagacccc gggacgccgc
     3601 cccggtagaa gctgacccca gatctgggcc cccgcggcat atgaccggtg aattct
//
