LOCUS       sequence-327560-        3662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-144(mutant2)~451 and miR-1-1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3662)
  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 3662)
  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-mir144m2~451-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..3662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1068..1149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1160..1177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1331..1919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1411..1430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2090..2881,2882..2950))
                     /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     2713..2732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2951..3055)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3088..3169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagggatc
      541 ctctagtcag ctgacgcgtg ctagcgcggc cgcgttctgc ccccagcccc tcacagtgct
      601 tttcaagcca tgcttcctgt gcccccagtg actacctggc tgggatatca tcatatactg
      661 taagtttgcg atgagacact acagtataga tgatgtacta gtccggtaag tcccagctct
      721 ggagcctgac aaggaggaca ggagagatgc tgcaagccca agaagctctc tgctcagcct
      781 gtcacaacct actgactgcc agggcacttg ggaatggcaa ggaaaccgtt accattactg
      841 agtttagtaa tggtaatggt tctcttgcta tacccagaaa acgtgccagg aagagaactc
      901 aggaccctga agcagactac tggaagggag actccaaagc ttgtcgacga tatctccaga
      961 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1021 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1081 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1141 tttcactgcc ccgagcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1201 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1501 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1741 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2041 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2101 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2161 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2221 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2281 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2341 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2401 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2461 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2521 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2581 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2641 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2701 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2761 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2821 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2881 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2941 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3001 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3061 ttccccgaaa agtgccacct gacgtcggca gtgaaaaaaa tgctttattt gtgaaatttg
     3121 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3181 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3241 aaacctctac aaatgtggta tggctgatta tgatcctcta gactgcaggg ctgtcggccg
     3301 cggcggcccc cgtggtgtgg agtgcccctc agtgcggcga gagaggtccc ggcctgagat
     3361 acatacttct ttacattcca tagcttagca ggtccatatg ggcatataaa gaagtatgtt
     3421 tcccaagcag gcagtctgca tgcagttcca agaccccggg acgccgcccc ggtagaagct
     3481 gaccccagat ctgggccccc gcggcatatg accggtgaat tctccaggcg atctgacggt
     3541 tcactaaacg agctctgctt atataggcct cccaccgtac acgccacctc gacatactcg
     3601 agtagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3661 cg
//
