LOCUS       sequence-328514-        3876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-144[309 nt]~451 and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3876)
  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 3876)
  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[309 nt]~451-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        391..594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     545..565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(873..891)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1368..1449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1460..1477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1631..2219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1711..1730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2390..3181,3182..3250))
                     /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     3013..3032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3251..3355)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3388..3469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gtacacgcca cctcgacata ctcgagtagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      421 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      481 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      541 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      601 gtgaaccgtc agatccgcta gggatcctct agtcagctga cgcgtgctag cgcggccgcg
      661 ttctgccccc agcccctcac agtgcttttc aagccatgct tcctgtgccc ccagtggggc
      721 cctggctggg atatcatcat atactgtaag tttgcgatga gacactacag tatagatgat
      781 gtactagtcc gggcaccccc agctctggag cctgacaagg aggacaggag agatgctgca
      841 agcccaaggg taccctggaa gatggaaccg ctggagagca actgcataag gctatgaaga
      901 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca
      961 cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc
     1021 tgaatacaaa tcacagaatc gtcgtatgca gtgacctgca ggaagctctc tgctcagcct
     1081 gtcacaacct actgactgcc agggcacttg ggaatggcaa ggaaaccgtt accattactg
     1141 agtttagtaa tggtaatggt tctcttgcta tacccagaaa acgtgccagg aagagaactc
     1201 aggaccctga agcagactac tggaagggag actccaaagc ttgtcgacga tatctccaga
     1261 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1321 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1381 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1441 tttcactgcc ccgagcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1501 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1561 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1621 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1681 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1741 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1801 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1861 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1921 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1981 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2041 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2101 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2161 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2221 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2281 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2341 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2401 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2461 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2521 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2581 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2641 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2701 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2761 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2821 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2881 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2941 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3001 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3061 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3121 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3181 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3241 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3301 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3361 ttccccgaaa agtgccacct gacgtcggca gtgaaaaaaa tgctttattt gtgaaatttg
     3421 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3481 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3541 aaacctctac aaatgtggta tggctgatta tgatcctcta gactgcaggg ctgtcggccg
     3601 cggcggcccc cgtggtgtgg agtgcccctc agtgcggcga gagaggtccc ggcctgagat
     3661 acatacttct ttacattcca tagcttagca ggtccatatg ggcatataaa gaagtatgtt
     3721 tcccaagcag gcagtctgca tgcagttcca agaccccggg acgccgcccc ggtagaagct
     3781 gaccccagat ctgggccccc gcggcatatg accggtgaat tctccaggcg atctgacggt
     3841 tcactaaacg agctctgctt atataggcct cccacc
//
