LOCUS       sequence-328520-        6798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express N-terminally truncated DroshaTN-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-D390TN(UniProt)-FLAG
FEATURES             Location/Qualifiers
     source          1..6798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        20..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        400..603
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     554..574
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     600..624
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          739..1564
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     CDS             4639..4662
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4735..4752)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4741..4948
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4957..5751)
                     /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(4957..5751)
                     /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(5068..5087)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5678..5697
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      6163..6751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6652..6671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg
       61 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      121 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      181 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      241 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
      301 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
      361 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      421 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      481 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
      541 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      601 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
      661 agaagacacc gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt
      721 ccccgtgcca agagtgacgt aagtaccgcc tatagactct ataggcacac ccctttggct
      781 cttatgcatg ctatactgtt tttggcttgg ggcctataca cccccgcttc cttatgctat
      841 aggtgatggt atagcttagc ctataggtgt gggttattga ccattattga ccactcccct
      901 attggtgacg atactttcca ttactaatcc ataacatggc tctttgccac aactatctct
      961 attggctata tgccaatact ctgtccttca gagactgaca cggactctgt atttttacag
     1021 gatggggtcc catttattat ttacaaattc acatatacaa caacgccgtc ccccgtgccc
     1081 gcagttttta ttaaacatag cgtgggatct ccacgcgaat ctcgggtacg tgttccggac
     1141 atgggctctt ctccggtagc ggcggagctt ccacatccga gccctggtcc catgcctcca
     1201 gcggctcatg gtcgctcggc agctccttgc tcctaacagt ggaggccaga cttaggcaca
     1261 gcacaatgcc caccaccacc agtgtgccgc acaaggccgt ggcggtaggg tatgtgtctg
     1321 aaaatgagct cggagattgg gctcgcaccg ctgacgcaga tggaagactt aaggcagcgg
     1381 cagaagaaga tgcaggcagc tgagttgttg tattctgata agagtcagag gtaactcccg
     1441 ttgcggtgct gttaacggtg gagggcagtg tagtctgagc agtactcgtt gctgccgcgc
     1501 gcgccaccag acataatagc tgacagacta acagactgtt cctttccatg ggtcttttct
     1561 gcagtcaccg tccttgacac gaagcttggt accgagctcg gatccactag tccagtgtgg
     1621 tggaattctg cagatatcca gcacagtggc ggccgcggga attcgattcg aggtaccatg
     1681 atgaaagagc ccgaggagac catgcctgac aagaatgagg aggaagaaga agaacttctt
     1741 aagcctgtgt ggattcgatg cactcattca gaaaactact actccagtga ccccatggat
     1801 caggtgggag attctacagt ggttggaacg agtaggcttc gtgacttata tgacaaattt
     1861 gaggaggagt tggggagcag gcaagaaaag gccaaagctg ctcggcctcc gtgggaacct
     1921 ccaaagacga agctcgatga agatttagag agttccagtg aatccgagtg tgagtctgat
     1981 gaggacagca cctgttctag cagctcagac tctgaagttt ttgacgttat tgcagaaatc
     2041 aaacgcaaaa aggcccaccc tgaccgactt catgatgaac tttggtacaa cgatccaggc
     2101 cagatgaatg atggaccact ctgcaaatgc agcgcaaagg caagacgcac aggaattagg
     2161 cacagcattt atcctggaga agaggccatc aagccctgtc gtcctatgac caacaatgct
     2221 ggcagacttt tccactaccg gatcacagtc tccccgccta cgaacttttt aactgacagg
     2281 ccaactgtta tagaatacga tgatcacgag tatatctttg aaggattttc tatgtttgca
     2341 catgcccccc tgaccaatat tccactgtgt aaagtaatta gattcaacat agactacacg
     2401 attcatttca ttgaagagat gatgccggag aatttttgtg tgaaagggct tgaactcttt
     2461 tcactgttcc tattcagaga tattttggaa ttatatgact ggaatcttaa aggtcctttg
     2521 tttgaagaca gccctccctg ctgcccaaga tttcatttca tgccacgttt tgtaagattt
     2581 cttccagatg gaggaaagga agtgctgtcc atgcaccaga ttctcctgta cttgttaagg
     2641 tgcagcaaag ccctggtgcc tgaggaggag attgccaata tgcttcagtg ggaggagctg
     2701 gagtggcaga aatatgcaga agaatgcaaa ggcatgattg ttaccaaccc tgggacgaaa
     2761 ccaagctctg tccgtatcga tcaactggat cgtgaacagt tcaaccccga tgtgattact
     2821 tttccgatta tcgtccactt tgggatacgc cctgcacagt tgagttatgc aggagaccca
     2881 cagtaccaaa aactgtggaa gagttatgtg aaacttcgcc acctcctagc aaatagtccc
     2941 aaagtcaaac aaactgacaa acagaagctg gcacagaggg aggaagccct ccaaaaaata
     3001 cggcagaaga atacaatgag acgagaagta acggtggagc taagtagcca aggattctgg
     3061 aaaactggca tccgttctga tgtctgtcag catgcaatga tgctacctgt tctgacccat
     3121 catatccgct accaccaatg cctaatgcat ttggacaagt tgataggata tactttccaa
     3181 gatcgttgtc tgttgcagct ggccatgact catccaagtc atcatttaaa ttttggaatg
     3241 aatcctgatc atgccaggaa ttcattatct aactgtggaa ttcggcagcc caaatacgga
     3301 gacagaaaag ttcatcacat gcacatgcgg aagaaaggga ttaacacctt gataaatatc
     3361 atgtcacgcc ttggccaaga tgacccaact ccctcgagga ttaaccacaa tgaacggttg
     3421 gaattcctgg gtgatgctgt tgttgaattt ctgaccagcg tccatttgta ctatttgttt
     3481 cctagtctgg aagaaggagg attagcaacc tatcggactg ccattgttca gaatcagcac
     3541 cttgccatgc tagcaaagaa acttgaactg gatcgattta tgctgtatgc tcacgggcct
     3601 gacctttgta gagaatcgga ccttcgacat gcaatggcca attgttttca agcgttaata
     3661 ggagctgttt acttggaggg aagcctggag gaagccaagc agttatttgg acgcttgctc
     3721 tttaatgatc cggacctgcg cgaagtctgg ctcaattatc ctctccaccc actccaacta
     3781 caagagccaa atactgatcg acaacttatt gaaacttctc cggttctaca aaaacttact
     3841 gagtttgaag aagcaattgg agtaattttt actcatgttc gacttctggc aagggcattc
     3901 acattgagaa ctgtgggatt taaccatctg accctaggcc acaatcagag aatggaattc
     3961 ctaggtgact ccataatgca actggtagcc acagagtact tattcattca tttcccagat
     4021 catcatgaag gacacttaac tttgttgcga agctctttgg tgaataatag aactcaggcc
     4081 aaggtagcgg aggagctggg catgcaggag tacgccataa ccaacgacaa gaccaagagg
     4141 cctgtggcgc ttcgcaccaa gaccttggcg gaccttttgc aatcatttat tgcagcgctg
     4201 tacattgata aggatttgga atatgttcat actttcatga atgtctgctt ctttccacga
     4261 ttgaaagagt tcattttgaa tcaggattgg aatgacccca aatcccagct tcagcagtgt
     4321 tgcttgacac ttaggacaga aggaaaagag ccagacattc ctctgtacaa gactctgcag
     4381 acagtgggcc catcccatgc ccgaacctac actgtggctg tttatttcaa gggagaaaga
     4441 ataggctgtg ggaaaggacc aagtattcag caagcggaaa tgggagcagc aatggatgcg
     4501 cttgaaaaat ataattttcc ccagatggcc catcagaagc ggttcatcga acggaagtac
     4561 agacaagagt taaaagaaat gaggtgggaa agagagcatc aagagagaga gccagatgag
     4621 actgaagaca tcaagaaaga ctacaaagac gatgacgata aataatctag agacaatcac
     4681 tagtgaattc gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     4741 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4801 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4861 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4921 gggaagacaa tagcaggcat gctggggagt cgaaattcag aagaactcgt caagaaggcg
     4981 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     5041 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     5101 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     5161 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     5221 gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     5281 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     5341 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     5401 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5461 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     5521 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     5581 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     5641 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5701 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     5761 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     5821 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     5881 caattccggt tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc
     5941 actgcaagct acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     6001 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg tgaaaaggat
     6061 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6481 aacggggggt tcgtgaacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6781 cctggccttt tgctggcc
//
