LOCUS       sequence-328516-        4476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-144[909 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 4476)
  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 4476)
  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[909 nt]~451-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..4476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        312..515
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     466..486
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(794..812)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1889..1970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1981..1998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2152..2740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2232..2251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2911..3702,3703..3771))
                     /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     3534..3553
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3772..3876)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3909..3990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      121 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      181 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      301 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      361 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      541 agggatcctc tagtcagctg acgcgtgcta gcgcggccgc gttctgcccc cagcccctca
      601 cagtgctttt caagccatgc ttcctgtgcc cccagtgggg ccctggctgg gatatcatca
      661 tatactgtaa gtttgcgatg agacactaca gtatagatga tgtactagtc cgggcacccc
      721 cagctctgga gcctgacaag gaggacagga gagatgctgc aagcccaagg gtaccctgga
      781 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
      841 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
      901 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
      961 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
     1021 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
     1081 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
     1141 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
     1201 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
     1261 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
     1321 ctctggatct actggtctgc ctaaaggtgt cgccctgcct catagaactg cctgcgtgag
     1381 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     1441 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     1501 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     1561 ggattacaag attcacctgc aggaagctct ctgctcagcc tgtcacaacc tactgactgc
     1621 cagggcactt gggaatggca aggaaaccgt taccattact gagtttagta atggtaatgg
     1681 ttctcttgct atacccagaa aacgtgccag gaagagaact caggaccctg aagcagacta
     1741 ctggaaggga gactccaaag cttgtcgacg atatctccag aggatcataa tcagccatac
     1801 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1861 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1921 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc cccgagcttc
     1981 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2041 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2161 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2221 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2281 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2341 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2401 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2461 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2521 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2581 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2641 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2701 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2761 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2821 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2881 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2941 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3001 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3061 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3121 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3181 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3241 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3301 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3361 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3421 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3481 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     3541 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3601 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3661 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3721 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3781 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3841 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3901 tgacgtcggc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3961 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4021 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4081 atggctgatt atgatcctct agactgcagg gctgtcggcc gcggcggccc ccgtggtgtg
     4141 gagtgcccct cagtgcggcg agagaggtcc cggcctgaga tacatacttc tttacattcc
     4201 atagcttagc aggtccatat gggcatataa agaagtatgt ttcccaagca ggcagtctgc
     4261 atgcagttcc aagaccccgg gacgccgccc cggtagaagc tgaccccaga tctgggcccc
     4321 cgcggcatat gaccggtgaa ttctccaggc gatctgacgg ttcactaaac gagctctgct
     4381 tatataggcc tcccaccgta cacgccacct cgacatactc gagtagttat taatagtaat
     4441 caattacggg gtcattagtt catagcccat atatgg
//
