LOCUS       sequence-328517-        7959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Drosha-FLAG in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  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 7959)
  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: pCK-Drosha(UniProt)-FLAG
FEATURES             Location/Qualifiers
     source          1..7959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1311..2136
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     primer_bind     complement(2251..2275)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2272..2475)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2301..2321)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2476..2855
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2922..3510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3002..3021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3922..4716
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3922..4716
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3976..3995)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4586..4605
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4725..4932
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4921..4938
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5011..5034)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cttgtcaggc atggtctcct cgggctcttt ttccttgatt gaggtatagt tcttgtcttt
       61 gccagaactc tggttgtcac tccaacggtc tttttcttcc tcccaacgag ctctcttctt
      121 ctccctactt ggggagcgat gattcacaat ctccaggggt ggggcccaag aatctgtatt
      181 tttaataatc tccccaggta attctggtgt gcatccagca ggttcaggaa caaccgataa
      241 accgtaactc cttccagatc tcttatactc ttttttgtag gacctttcca gagatggtga
      301 tcttcggttg tctcgatgcc tgtgtctctc ccgttctcgc tctctgctcc gttcgtagct
      361 gcggtggcga gatggtgttc tccctcggtc ataatcagat ctgtaccggc tgtcttgtct
      421 tctcctgtcg ggactgcggc ctcgctcccg ccgatccagg gaccgatgcc tctcacctcg
      481 cccatgactg tgatctcggt gcctgtggtc atcatagtgt ttcagccttt ctggggacct
      541 tctctcactg ggagcctttg ggagtgggta tggagggaga tgtctgaaat gaggactact
      601 gctgttatta gcactgggca ggaaagaact agggttgttc tggaaactat taaaactggg
      661 aggtgggaag ttgtggtgag aatagcccgg agggtactga taattaacct gctgcggcat
      721 gactggaggg ggcgggggat gaggcatgga gggagggggc atcatgaagg ggaaagtgcc
      781 ttgtccagga ggtgccccag ggactggggg gttattagga caaggcattg gtggtggcat
      841 gggaggaaaa caaggaggaa ctgggaaggg gtgcctcatc tggtggttgg ggaaaggcgg
      901 cctgattggg caggggggaa gagggccttg cgctgacgga ggcatgggtg gggggaaggg
      961 tacaaagtct ggtcgtggag ggagaaaatt gggggctgga gagtttgaga aagtggtgga
     1021 aggggcactt ggaggttcat attgatattg cacaggaggc tgctgagggt gaagcagcct
     1081 cagattttgg ggcctaaagg atggtgctga gggtctggct ccatgtcctc ctcgtcctcg
     1141 gggacaccct cgtcccgggt ggaacgacat tctgtgacat gtgtttccct gcatcatggt
     1201 acctcgaatc gaattcccgc ggccgccact gtgctggata tctgcagaat tccaccacac
     1261 tggactagtg gatccgagct cggtaccaag cttcgtgtca aggacggtga ctgcagaaaa
     1321 gacccatgga aaggaacagt ctgttagtct gtcagctatt atgtctggtg gcgcgcgcgg
     1381 cagcaacgag tactgctcag actacactgc cctccaccgt taacagcacc gcaacgggag
     1441 ttacctctga ctcttatcag aatacaacaa ctcagctgcc tgcatcttct tctgccgctg
     1501 ccttaagtct tccatctgcg tcagcggtgc gagcccaatc tccgagctca ttttcagaca
     1561 cataccctac cgccacggcc ttgtgcggca cactggtggt ggtgggcatt gtgctgtgcc
     1621 taagtctggc ctccactgtt aggagcaagg agctgccgag cgaccatgag ccgctggagg
     1681 catgggacca gggctcggat gtggaagctc cgccgctacc ggagaagagc ccatgtccgg
     1741 aacacgtacc cgagattcgc gtggagatcc cacgctatgt ttaataaaaa ctgcgggcac
     1801 gggggacggc gttgttgtat atgtgaattt gtaaataata aatgggaccc catcctgtaa
     1861 aaatacagag tccgtgtcag tctctgaagg acagagtatt ggcatatagc caatagagat
     1921 agttgtggca aagagccatg ttatggatta gtaatggaaa gtatcgtcac caatagggga
     1981 gtggtcaata atggtcaata acccacacct ataggctaag ctataccatc acctatagca
     2041 taaggaagcg ggggtgtata ggccccaagc caaaaacagt atagcatgca taagagccaa
     2101 aggggtgtgc ctatagagtc tataggcggt acttacgtca ctcttggcac ggggaatccg
     2161 cgttccaatg caccgttccc ggccgcggag gctggatcgg tcccggtgtc ttctatggag
     2221 gtcaaaacag cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctctgct
     2281 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaacgggg cggggttatt
     2341 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2401 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tggtgtactg
     2461 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2521 aaagtcccgt aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2581 gtcaataggg ggcggacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     2641 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     2701 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     2761 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     2821 tgattactat taataactag tcaataatca atgtcaacgc gcatgtgagc aaaaggccag
     2881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3181 gaaccccccg ttcagcccga ccgctgtgcc ttatccggta actatcgtct tgagtccaac
     3241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3601 atcttcacct agatcctttt cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     3661 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag
     3721 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc
     3781 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     3841 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag
     3901 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3961 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4021 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4081 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4141 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4201 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4261 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4321 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4381 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4441 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4501 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4561 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4621 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4681 atcgccttct atcgccttct tgacgagttc ttctgaattt cgactcccca gcatgcctgc
     4741 tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat
     4801 gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag
     4861 tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac
     4921 tagaaggcac agtcgaggct gatcagcggg tttaaacggg ccctctagac tcgagcggcc
     4981 gcgaattcac tagtgattgt ctctagatta tttatcgtca tcgtctttgt agtctttctt
     5041 gatgtcttca gtctcatctg gctctctctc ttgatgctct ctttcccacc tcatttcttt
     5101 taactcttgt ctgtacttcc gttcgatgaa ccgcttctga tgggccatct ggggaaaatt
     5161 atatttttca agcgcatcca ttgctgctcc catttccgct tgctgaatac ttggtccttt
     5221 cccacagcct attctttctc ccttgaaata aacagccaca gtgtaggttc gggcatggga
     5281 tgggcccact gtctgcagag tcttgtacag aggaatgtct ggctcttttc cttctgtcct
     5341 aagtgtcaag caacactgct gaagctggga tttggggtca ttccaatcct gattcaaaat
     5401 gaactctttc aatcgtggaa agaagcagac attcatgaaa gtatgaacat attccaaatc
     5461 cttatcaatg tacagcgctg caataaatga ttccaaaagg tccgccaagg tcttggtgcg
     5521 aagcgccaca ggcctcttgg tcttgtcgtt ggttatggcg tactcctgca tgcccagctc
     5581 ctccgctacc ttggcctgag ttctattatt caccaaagag cttcgcaaca aagttaagtg
     5641 tccttcatga tgatctggga aatgaatgaa taagtactct gtggctacca gttgcattat
     5701 ggagtcacct aggaattcca ttctctgatt gtggcctagg gtcagatggt taaatcccac
     5761 agttctcaat gtgaatgccc ttgccagaag tcgaacatga gtaaaaatta ctccaattgc
     5821 ttcttcaaac tcagtaagtt tttgtagaac cggagaagtt tcaataagtt gtcgatcagt
     5881 atttggctct tgtagttgga gtgggtggag aggataattg agccagactt cgcgcaggtc
     5941 cggatcatta aagagcaagc gtccaaataa ctgcttggct tcctccaggc ttccctccaa
     6001 gtaaacagct cctattaacg cttcaaaaca attggccatt gcatgtcgaa ggtccgattc
     6061 tctacaaagg tcaggcccgt gagcatacag cataaatcga tccagttcaa gtttctttgc
     6121 tagcatggca aggtgctgat tctgaacaat ggcagtccga taggttgcta atcctccttc
     6181 ttccagacta ggaaacaaat agtacaaatg gacgctggtc agaaattcaa caacagcatc
     6241 acccaggaat tccaaccgtt cattgtggtt aatcctcgag ggagttgggt catcttggcc
     6301 aaggcgtgac atgatattta tcaaggtgtt aatccctttc ttccgcatgt gcatgtgatg
     6361 aacttttctg tctccgtatt tgggctgccg aattccacag ttagataatg aattcctggc
     6421 atgatcagga ttcattccaa aatttaaatg atgacttgga tgagtcatgg ccagctgcaa
     6481 cagacaacga tcttggaaag tatatcctat caacttgtcc aaatgcatta ggcattggtg
     6541 gtagcggata tgatgggtca gaacaggtag catcattgca tgctgacaga catcagaacg
     6601 gatgccagtt ttccagaatc cttggctact tagctccacc gttacttctc gtctcattgt
     6661 attcttctgc cgtatttttt ggagggcttc ctccctctgt gccagcttct gtttgtcagt
     6721 ttgtttgact ttgggactat ttgctaggag gtggcgaagt ttcacataac tcttccacag
     6781 tttttggtac tgtgggtctc ctgcataact caactgtgca gggcgtatcc caaagtggac
     6841 gataatcgga aaagtaatca catcggggtt gaactgttca cgatccagtt gatcgatacg
     6901 gacagagctt ggtttcgtcc cagggttggt aacaatcatg cctttgcatt cttctgcata
     6961 tttctgccac tccagctcct cccactgaag catattggca atctcctcct caggcaccag
     7021 ggctttgctg caccttaaca agtacaggag aatctggtgc atggacagca cttcctttcc
     7081 tccatctgga agaaatctta caaaacgtgg catgaaatga aatcttgggc agcagggagg
     7141 gctgtcttca aacaaaggac ctttaagatt ccagtcatat aattccaaaa tatctctgaa
     7201 taggaacagt gaaaagagtt caagcccttt cacacaaaaa ttctccggca tcatctcttc
     7261 aatgaaatga atcgtgtagt ctatgttgaa tctaattact ttacacagtg gaatattggt
     7321 caggggggca tgtgcaaaca tagaaaatcc ttcaaagata tactcgtgat catcgtattc
     7381 tataacagtt ggcctgtcag ttaaaaagtt cgtaggcggg gagactgtga tccggtagtg
     7441 gaaaagtctg ccagcattgt tggtcatagg acgacagggc ttgatggcct cttctccagg
     7501 ataaatgctg tgcctaattc ctgtgcgtct tgcctttgcg ctgcatttgc agagtggtcc
     7561 atcattcatc tggcctggat cgttgtacca aagttcatca tgaagtcggt cagggtgggc
     7621 ctttttgcgt ttgatttctg caataacgtc aaaaacttca gagtctgagc tgctagaaca
     7681 ggtgctgtcc tcatcagact cacactcgga ttcactggaa ctctctaaat cttcatcgag
     7741 cttcgtcttt ggaggttccc acggaggccg agcagctttg gccttttctt gcctgctccc
     7801 caactcctcc tcaaatttgt catataagtc acgaagccta ctcgttccaa ccactgtaga
     7861 atctcccacc tgatccatgg ggtcactgga gtagtagttt tctgaatgag tgcatcgaat
     7921 ccacacaggc ttaagaagtt cttcttcttc ctcctcatt
//
