LOCUS       sequence-328519-        6798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-terminally truncated Drosha-FLAG in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6798)
  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 6798)
  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-D390(UniProt)-FLAG
FEATURES             Location/Qualifiers
     source          1..6798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        32..411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     566..586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     612..636
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          751..1576
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     CDS             4651..4674
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4747..4764)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4753..4960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4969..5763)
                     /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             complement(4969..5763)
                     /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(5080..5099)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5690..5709
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      6175..6763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6664..6683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctgg ccttttgctc acatgcgcgt tgacattgat tattgactag ttattaatag
       61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
      301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
      361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      421 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      541 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      601 tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
      661 tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccggga acggtgcatt
      721 ggaacgcgga ttccccgtgc caagagtgac gtaagtaccg cctatagact ctataggcac
      781 acccctttgg ctcttatgca tgctatactg tttttggctt ggggcctata cacccccgct
      841 tccttatgct ataggtgatg gtatagctta gcctataggt gtgggttatt gaccattatt
      901 gaccactccc ctattggtga cgatactttc cattactaat ccataacatg gctctttgcc
      961 acaactatct ctattggcta tatgccaata ctctgtcctt cagagactga cacggactct
     1021 gtatttttac aggatggggt cccatttatt atttacaaat tcacatatac aacaacgccg
     1081 tcccccgtgc ccgcagtttt tattaaacat agcgtgggat ctccacgcga atctcgggta
     1141 cgtgttccgg acatgggctc ttctccggta gcggcggagc ttccacatcc gagccctggt
     1201 cccatgcctc cagcggctca tggtcgctcg gcagctcctt gctcctaaca gtggaggcca
     1261 gacttaggca cagcacaatg cccaccacca ccagtgtgcc gcacaaggcc gtggcggtag
     1321 ggtatgtgtc tgaaaatgag ctcggagatt gggctcgcac cgctgacgca gatggaagac
     1381 ttaaggcagc ggcagaagaa gatgcaggca gctgagttgt tgtattctga taagagtcag
     1441 aggtaactcc cgttgcggtg ctgttaacgg tggagggcag tgtagtctga gcagtactcg
     1501 ttgctgccgc gcgcgccacc agacataata gctgacagac taacagactg ttcctttcca
     1561 tgggtctttt ctgcagtcac cgtccttgac acgaagcttg gtaccgagct cggatccact
     1621 agtccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgcgg gaattcgatt
     1681 cgaggtacca tgatgaaaga gcccgaggag accatgcctg acaagaatga ggaggaagaa
     1741 gaagaacttc ttaagcctgt gtggattcga tgcactcatt cagaaaacta ctactccagt
     1801 gaccccatgg atcaggtggg agattctaca gtggttggaa cgagtaggct tcgtgactta
     1861 tatgacaaat ttgaggagga gttggggagc aggcaagaaa aggccaaagc tgctcggcct
     1921 ccgtgggaac ctccaaagac gaagctcgat gaagatttag agagttccag tgaatccgag
     1981 tgtgagtctg atgaggacag cacctgttct agcagctcag actctgaagt ttttgacgtt
     2041 attgcagaaa tcaaacgcaa aaaggcccac cctgaccgac ttcatgatga actttggtac
     2101 aacgatccag gccagatgaa tgatggacca ctctgcaaat gcagcgcaaa ggcaagacgc
     2161 acaggaatta ggcacagcat ttatcctgga gaagaggcca tcaagccctg tcgtcctatg
     2221 accaacaatg ctggcagact tttccactac cggatcacag tctccccgcc tacgaacttt
     2281 ttaactgaca ggccaactgt tatagaatac gatgatcacg agtatatctt tgaaggattt
     2341 tctatgtttg cacatgcccc cctgaccaat attccactgt gtaaagtaat tagattcaac
     2401 atagactaca cgattcattt cattgaagag atgatgccgg agaatttttg tgtgaaaggg
     2461 cttgaactct tttcactgtt cctattcaga gatattttgg aattatatga ctggaatctt
     2521 aaaggtcctt tgtttgaaga cagccctccc tgctgcccaa gatttcattt catgccacgt
     2581 tttgtaagat ttcttccaga tggaggaaag gaagtgctgt ccatgcacca gattctcctg
     2641 tacttgttaa ggtgcagcaa agccctggtg cctgaggagg agattgccaa tatgcttcag
     2701 tgggaggagc tggagtggca gaaatatgca gaagaatgca aaggcatgat tgttaccaac
     2761 cctgggacga aaccaagctc tgtccgtatc gatcaactgg atcgtgaaca gttcaacccc
     2821 gatgtgatta cttttccgat tatcgtccac tttgggatac gccctgcaca gttgagttat
     2881 gcaggagacc cacagtacca aaaactgtgg aagagttatg tgaaacttcg ccacctccta
     2941 gcaaatagtc ccaaagtcaa acaaactgac aaacagaagc tggcacagag ggaggaagcc
     3001 ctccaaaaaa tacggcagaa gaatacaatg agacgagaag taacggtgga gctaagtagc
     3061 caaggattct ggaaaactgg catccgttct gatgtctgtc agcatgcaat gatgctacct
     3121 gttctgaccc atcatatccg ctaccaccaa tgcctaatgc atttggacaa gttgatagga
     3181 tatactttcc aagatcgttg tctgttgcag ctggccatga ctcatccaag tcatcattta
     3241 aattttggaa tgaatcctga tcatgccagg aattcattat ctaactgtgg aattcggcag
     3301 cccaaatacg gagacagaaa agttcatcac atgcacatgc ggaagaaagg gattaacacc
     3361 ttgataaata tcatgtcacg ccttggccaa gatgacccaa ctccctcgag gattaaccac
     3421 aatgaacggt tggaattcct gggtgatgct gttgttgaat ttctgaccag cgtccatttg
     3481 tactatttgt ttcctagtct ggaagaagga ggattagcaa cctatcggac tgccattgtt
     3541 cagaatcagc accttgccat gctagcaaag aaacttgaac tggatcgatt tatgctgtat
     3601 gctcacgggc ctgacctttg tagagaatcg gaccttcgac atgcaatggc caattgtttt
     3661 gaagcgttaa taggagctgt ttacttggag ggaagcctgg aggaagccaa gcagttattt
     3721 ggacgcttgc tctttaatga tccggacctg cgcgaagtct ggctcaatta tcctctccac
     3781 ccactccaac tacaagagcc aaatactgat cgacaactta ttgaaacttc tccggttcta
     3841 caaaaactta ctgagtttga agaagcaatt ggagtaattt ttactcatgt tcgacttctg
     3901 gcaagggcat tcacattgag aactgtggga tttaaccatc tgaccctagg ccacaatcag
     3961 agaatggaat tcctaggtga ctccataatg caactggtag ccacagagta cttattcatt
     4021 catttcccag atcatcatga aggacactta actttgttgc gaagctcttt ggtgaataat
     4081 agaactcagg ccaaggtagc ggaggagctg ggcatgcagg agtacgccat aaccaacgac
     4141 aagaccaaga ggcctgtggc gcttcgcacc aagaccttgg cggacctttt ggaatcattt
     4201 attgcagcgc tgtacattga taaggatttg gaatatgttc atactttcat gaatgtctgc
     4261 ttctttccac gattgaaaga gttcattttg aatcaggatt ggaatgaccc caaatcccag
     4321 cttcagcagt gttgcttgac acttaggaca gaaggaaaag agccagacat tcctctgtac
     4381 aagactctgc agacagtggg cccatcccat gcccgaacct acactgtggc tgtttatttc
     4441 aagggagaaa gaataggctg tgggaaagga ccaagtattc agcaagcgga aatgggagca
     4501 gcaatggatg cgcttgaaaa atataatttt ccccagatgg cccatcagaa gcggttcatc
     4561 gaacggaagt acagacaaga gttaaaagaa atgaggtggg aaagagagca tcaagagaga
     4621 gagccagatg agactgaaga catcaagaaa gactacaaag acgatgacga taaataatct
     4681 agagacaatc actagtgaat tcgcggccgc tcgagtctag agggcccgtt taaacccgct
     4741 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     4801 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4861 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4921 agggggagga ttgggaagac aatagcaggc atgctgggga gtcgaaattc agaagaactc
     4981 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     5041 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     5101 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     5161 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     5221 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     5281 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     5341 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     5401 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     5461 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     5521 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     5581 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     5641 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     5701 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     5761 catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat
     5821 ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
     5881 cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg
     5941 ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca
     6001 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     6061 cgtgaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6121 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6181 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6241 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6301 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6361 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6421 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcaca
     6481 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6541 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6601 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6661 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6721 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6781 ctttttacgg ttcctggc
//
