LOCUS       sequence-327564-        6639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miR-144[3072 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 6639)
  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 6639)
  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[3072 nt]~451-CMV2-mir1
FEATURES             Location/Qualifiers
     source          1..6639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    442..523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(534..551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(705..1293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(785..804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1464..2255,2256..2324))
                     /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     2087..2106
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2325..2429)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2462..2543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        3037..3340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3341..3544
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3495..3515
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 accaaaacca caacgtgctg ttaaaaggta gagtggcatg tgaaattaat tctggttatg
       61 agttagtttt aactgaatta attttagaca actttttagg aagctttgaa cctgaagaga
      121 ttgtagcccc tgcaggaagc tctctgctca gcctgtcaca acctactgac tgccagggca
      181 cttgggaatg gcaaggaaac cgttaccatt actgagttta gtaatggtaa tggttctctt
      241 gctataccca gaaaacgtgc caggaagaga actcaggacc ctgaagcaga ctactggaag
      301 ggagactcca aagcttgtcg acgatatctc cagaggatca taatcagcca taccacattt
      361 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
      421 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
      481 aatagcatca caaatttcac aaataaagca tttttttcac tgccccgagc ttcctcgctc
      541 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      601 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      661 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      721 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      781 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      841 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      901 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      961 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1021 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1081 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1141 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1201 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1261 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1321 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1381 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1441 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1501 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1561 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1621 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1681 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1741 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1801 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1861 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1921 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1981 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2041 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     2101 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2161 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2221 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2281 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2341 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2401 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     2461 ggcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2521 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2581 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     2641 attatgatcc tctagactgc agggctgtcg gccgcggcgg cccccgtggt gtggagtgcc
     2701 cctcagtgcg gcgagagagg tcccggcctg agatacatac ttctttacat tccatagctt
     2761 agcaggtcca tatgggcata taaagaagta tgtttcccaa gcaggcagtc tgcatgcagt
     2821 tccaagaccc cgggacgccg ccccggtaga agctgacccc agatctgggc ccccgcggca
     2881 tatgaccggt gaattctcca ggcgatctga cggttcacta aacgagctct gcttatatag
     2941 gcctcccacc gtacacgcca cctcgacata ctcgagtagt tattaatagt aatcaattac
     3001 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3061 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3121 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3181 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3241 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     3301 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3361 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3421 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     3481 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     3541 agctggttta gtgaaccgtc agatccgcta gggatcctct agtcagctga cgcgtgctag
     3601 cgcggccgcg ttctgccccc agcccctcac agtgcttttc aagccatgct tcctgtgccc
     3661 ccagtggggc cctggctggg atatcatcat atactgtaag tttgcgatga gacactacag
     3721 tatagatgat gtactagtcc gggcaccccc agctctggag cctgacaagg aggacaggag
     3781 agatgctgca agcccaaggg taccgaattc ccatgaaatc tgtcaagaca gattcagaac
     3841 aaaatggatc ttctacaatg gccaacgcta ctaaattgtt acacaaggat ggtcaaggtt
     3901 tattcgatat tccagagggt atgaatagag gcattaagcc gatggattct ccagctgaaa
     3961 atgaggatca aaacggacag ttcaaagaat tgaaacagct aaatgagata gataatgaat
     4021 tagacataag gattgaggcg aacgaggcaa agttaaaaga ggaggaaaaa tctgccaagt
     4081 caatttctga agaaattatg gaggaagcca ctgaagaaac gactgccgat aatgctgatg
     4141 atgccgaaat cgatgaacta ttaccaattg gaatagactt tggcagaact aaaccagttt
     4201 cgaaaagtgt tcctgttaaa aaggaatggg cccatgttgt tgatttgaac cataagatcg
     4261 aaaatttcga tgagttgatt ccaaaccccg ctagatcgtg gccttttgaa ttggatacat
     4321 ttcaaaagga agccgtctat catctcgaac aaggtgattc cgtatttgtt gccgcccaca
     4381 catctgctgg taaaactgtt gttgcagaat acgcaattgc catggcacat agaaacatga
     4441 ctaaaactat atatacatca cccattaaag ccttatccaa tcaaaaattc cgagacttta
     4501 aagaaacttt tgacgatgtt aatattgggt tgattactgg tgatgtgcaa attaatccgg
     4561 atgctaactg tttgattatg acgaccgaaa ttttaagatc aatgctttat agaggtgccg
     4621 atttgatcag agatgtagag tttgtcattt tcgatgaagt tcactacgtt aatgatcaag
     4681 accgtggtgt tgtctgggaa gaagttatta ttatgcttcc acagcatgtc aagtttatct
     4741 tattatctgc caccgttcct aacacttatg agtttgctaa ttggattgga agaactaagc
     4801 aaaaaaatat ttatgtcatt tccacaccaa aaagacccgt tccattggaa ataaatatat
     4861 gggctaaaaa ggaactgata ccggttatta atcaaaattc agaattttta gaagctaatt
     4921 tcaggaaaca taaagagatt ctaaatgggg agagtgcaaa aggggctcct tcaaaaactg
     4981 acaacgggag aggtggatcc acagcaagag ggggccgtgg tggatccaat actcgagatg
     5041 gaagaggagg ccgtggtaat tcaacacgtg gaggtgccaa ccgtggaggt tcaagaggtg
     5101 ctggcgcaat tggctccaac aagcgtaagt ttttcactca agatggtcct tcgaaaaaaa
     5161 catggcccga aattgtcaat tacctaagga aaagagagct tctaccaatg gttgttttcg
     5221 tatttagtaa aaagaggtgc gaggaatatg ctgactggtt ggagggaatt aacttctgca
     5281 acaataagga aaaatcacaa atccacatgt ttattgagaa gtcgattact cgtttgaaaa
     5341 aagaggatag agatttgcct caaattctta aaaccagatc attacttgaa cgtggtatag
     5401 ctgttcatca tggggggcta ttgcccattg tcaaggagtt gatcgaaata ttattttcca
     5461 agggttttat caaagtttta tttgcaacag aaacttttgc catgggtttg aaccttccca
     5521 caagaacagt tatctttagc agcatccgta aacatgatgg taatggatta agagaactga
     5581 ctccaggtga gtttacccaa atggctggta gagctggtag aaggggcttg gattccactg
     5641 gtacagtcat tgtgatggca tataatagtc ctttatctat tgctacgttc aaagaagtaa
     5701 cgatgggtgt tccgacgagg ttgcagtctc agtttagatt aacatacaat atgatattaa
     5761 atcttttaag aattgaggcg ttgagagttg aagaaatgat caagtattct tttagtgaga
     5821 atgccaaaga aactttacaa cctgaacatg aaaaacaaat caaagtatta caagaggaat
     5881 tacaaaccat agagtacaaa agttgtgaaa tctgtgataa tgatatcgaa aagtttttgg
     5941 aattgatgct ggcatataag gaggcaacag tcaacctaat gcaagaaatg gttaaatcgc
     6001 cttcgatatt gcatatcttg aaagagggca gactcgttgc ttttagggac cccaatgatt
     6061 gcttgaaatt aggatttgta tttaaagttt ctctgaagga tgctgtttgt gtcattatga
     6121 cgttcacaaa accatacaaa ctcccaaatg gtgagccaaa tcatttaata tactttccaa
     6181 aagcggatgg atacagaaga aggaacttcc ctaaattcca gaagactgat ttttatatgg
     6241 aagaagttcc tgtgacggct attgaagtga tcactaaacg caagtttgcc gcacccttgg
     6301 gaaaagtgat taaaaaggat gttgccgctt tgaacgagtt taacgctgaa acaaataaca
     6361 ttttggacgg caaaacgcta aaagaagcaa ttaacattga aaagcaaggt ttaaaaattc
     6421 accaaatatt gttagaccgt acaaatataa gggatgagat attcaaacta aaaagcatta
     6481 aatgtcccaa cctaagccaa cacattgttc ccaagtttaa ggcgcatgta attaaaaaga
     6541 aaattgaaga attgtaccat ttgatgtctg accaaaatct gagcttacta cccgattatg
     6601 aaaagagatt ggctgttctg aaagacactg aatttatcg
//
