LOCUS       sequence-340168-        7959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mutant human Drosha protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  AUTHORS   Zhang X, Li P, Lin J, Huang H, Yin B, Zeng Y
  TITLE     The insertion in the double-stranded RNA binding domain of human 
            Drosha is important for its function.
  JOURNAL   Biochim Biophys Acta Gene Regul Mech. 2017 Dec;1860(12):1179-1188. 
            doi: 10.1016/j.bbagrm.2017.10.004. Epub 2017 Nov 3.
  PUBMED    29109067
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.bbagrm.2017.10.004"; journalName: "Biochim Biophys Acta 
            Gene Regul Mech"; date: "2017-12"; volume: "1860"; issue: "12"; 
            pages: "1179-1188"
COMMENT     Sequence Label: pCK-Drosha-FLAG(delta6)
FEATURES             Location/Qualifiers
     source          1..7959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          769..1594
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     CDS             5830..5853
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5926..5943)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5932..6139
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(6148..6942)
                     /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(6148..6942)
                     /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(6259..6278)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6869..6888
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      7354..7942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7843..7862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgcgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
      721 ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc
      781 tatagactct ataggcacac ccctttggct cttatgcatg ctatactgtt tttggcttgg
      841 ggcctataca cccccgcttc cttatgctat aggtgatggt atagcttagc ctataggtgt
      901 gggttattga ccattattga ccactcccct attggtgacg atactttcca ttactaatcc
      961 ataacatggc tctttgccac aactatctct attggctata tgccaatact ctgtccttca
     1021 gagactgaca cggactctgt atttttacag gatggggtcc catttattat ttacaaattc
     1081 acatatacaa caacgccgtc ccccgtgccc gcagttttta ttaaacatag cgtgggatct
     1141 ccacgcgaat ctcgggtacg tgttccggac atgggctctt ctccggtagc ggcggagctt
     1201 ccacatccga gccctggtcc catgcctcca gcggctcatg gtcgctcggc agctccttgc
     1261 tcctaacagt ggaggccaga cttaggcaca gcacaatgcc caccaccacc agtgtgccgc
     1321 acaaggccgt ggcggtaggg tatgtgtctg aaaatgagct cggagattgg gctcgcaccg
     1381 ctgacgcaga tggaagactt aaggcagcgg cagaagaaga tgcaggcagc tgagttgttg
     1441 tattctgata agagtcagag gtaactcccg ttgcggtgct gttaacggtg gagggcagtg
     1501 tagtctgagc agtactcgtt gctgccgcgc gcgccaccag acataatagc tgacagacta
     1561 acagactgtt cctttccatg ggtcttttct gcagtcaccg tccttgacac gaagcttggt
     1621 accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca gcacagtggc
     1681 ggccgcggga attcgattcg aggtaccatg atgcagggaa acacatgtca cagaatgtcg
     1741 ttccacccgg gacgagggtg tccccgagga cgaggaggac atggagccag acccccagca
     1801 ccatccttta ggccccaaaa tctgaggctg cttcaccctc agcagcctcc tgtgcaatat
     1861 caatatgaac ctccaagtgc cccttccacc actttctcaa actctccagc ccccaatttt
     1921 ctccctccac gaccagactt tgtacccttc cccccaccca tgcctccgtc agcgcaaggc
     1981 cctcttcccc cctgcccaat caggccgcct ttccccaacc accagatgag gcaccccttc
     2041 ccagttcctc cttgttttcc tcccatgcca ccaccaatgc cttgtcctaa taacccccca
     2101 gtccctgggg tacctcctgg acaaggcact ttccccttca tgatgccccc tccctccatg
     2161 cctcatcccc cgccccctcc agtcatgccg cagcaggtta attatcagta ccctccgggc
     2221 tattctcacc acaacttccc acctcccagt tttaatagtt tccagaacaa ccctagttct
     2281 ttcctgccca gtgctaataa cagcggtagt cctcatttca gacatctccc tccataccca
     2341 ctcccaaagg ctcccagtga gagaaggtcc ccagaaaggc tgaaacacta tgatgaccac
     2401 aggcaccgag atcacagtca tgggcgaggt gagaggcatc ggtccctgga tcggcgggag
     2461 cgaggccgca gtcccgacag gagaagacaa gacagccggt acagatctga ttatgaccga
     2521 gggagaacac catctcgcca ccgcagctac gaacggagca gagagcgaga acgggagaga
     2581 cacaggcatc gagacaaccg aagatcacca tctctggaaa ggtcctacaa aaaagagtat
     2641 aagagatctg gaaggagtta cggtttattg gttgttcctg aacctgctgg atgcacacca
     2701 gaattacctg gggagattat taaaaataca gattcttggg ccccacccct ggagattgtg
     2761 aatcatcgct ccccaagtag ggagaagaag agagctcgtt gggaggaaga aaaagaccgt
     2821 tggagtgaca accagagttc tggcaaagac aagaactata cctcaatcaa ggaaaaagag
     2881 cccgaggaga ccatgcctga caagaatgag gaggaagaag aagaacttct taagcctgtg
     2941 tggattcgat gcactcattc agaaaactac tactccagtg accccatgga tcaggtggga
     3001 gattctacag tggttggaac gagtaggctt cgtgacttat atgacaaatt tgaggaggag
     3061 ttggggagca ggcaagaaaa ggccaaagct gctcggcctc cgtgggaacc tccaaagacg
     3121 aagctcgatg aagatttaga gagttccagt gaatccgagt gtgagtctga tgaggacagc
     3181 acctgttcta gcagctcaga ctctgaagtt tttgacgtta ttgcagaaat caaacgcaaa
     3241 aaggcccacc ctgaccgact tcatgatgaa ctttggtaca acgatccagg ccagatgaat
     3301 gatggaccac tctgcaaatg cagcgcaaag gcaagacgca caggaattag gcacagcatt
     3361 tatcctggag aagaggccat caagccctgt cgtcctatga ccaacaatgc tggcagactt
     3421 ttccactacc ggatcacagt ctccccgcct acgaactttt taactgacag gccaactgtt
     3481 atagaatacg atgatcacga gtatatcttt gaaggatttt ctatgtttgc acatgccccc
     3541 ctgaccaata ttccactgtg taaagtaatt agattcaaca tagactacac gattcatttc
     3601 attgaagaga tgatgccgga gaatttttgt gtgaaagggc ttgaactctt ttcactgttc
     3661 ctattcagag atattttgga attatatgac tggaatctta aaggtccttt gtttgaagac
     3721 agccctccct gctgcccaag atttcatttc atgccacgtt ttgtaagatt tcttccagat
     3781 ggaggaaagg aagtgctgtc catgcaccag attctcctgt acttgttaag gtgcagcaaa
     3841 gccctggtgc ctgaggagga gattgccaat atgcttcagt gggaggagct ggagtggcag
     3901 aaatatgcag aagaatgcaa aggcatgatt gttaccaacc ctgggacgaa accaagctct
     3961 gtccgtatcg atcaactgga tcgtgaacag ttcaaccccg atgtgattac ttttccgatt
     4021 atcgtccact ttgggatacg ccctgcacag ttgagttatg caggagaccc acagtaccaa
     4081 aaactgtgga agagttatgt gaaacttcgc cacctcctag caaatagtcc caaagtcaaa
     4141 caaactgaca aacagaagct ggcacagagg gaggaagccc tccaaaaaat acggcagaag
     4201 aatacaatga gacgagaagt aacggtggag ctaagtagcc aaggattctg gaaaactggc
     4261 atccgttctg atgtctgtca gcatgcaatg atgctacctg ttctgaccca tcatatccgc
     4321 taccaccaat gcctaatgca tttggacaag ttgataggat atactttcca agatcgttgt
     4381 ctgttgcagc tggccatgac tcatccaagt catcatttaa attttggaat gaatcctgat
     4441 catgccagga attcattatc taactgtgga attcggcagc ccaaatacgg agacagaaaa
     4501 gttcatcaca tgcacatgcg gaagaaaggg attaacacct tgataaatat catgtcacgc
     4561 cttggccaag atgacccaac tccctcgagg attaaccaca atgaacggtt ggaattcctg
     4621 ggtgatgctg ttgttgaatt tctgaccagc gtccatttgt actatttgtt tcctagtctg
     4681 gaagaaggag gattagcaac ctatcggact gccattgttc agaatcagca ccttgccatg
     4741 ctagcaaaga aacttgaact ggatcgattt atgctgtatg ctcacgggcc tgacctttgt
     4801 agagaatcgg accttcgaca tgcaatggcc aattgttttg aagcgttaat aggagctgtt
     4861 tacttggagg gaagcctgga ggaagccaag cagttatttg gacgcttgct ctttaatgat
     4921 ccggacctgc gcgaagtctg gctcaattat cctctccacc cactccaact acaagagcca
     4981 aatactgatc gacaacttat tgaaacttct ccggttctac aaaaacttac tgagtttgaa
     5041 gaagcaattg gagtaatttt tactcatgtt cgacttctgg caagggcatt cacattgaga
     5101 actgtgggat ttaaccatct gaccctaggc cacaatcaga gaatgaaatt cctaggtgac
     5161 tccataatgc aactggtagc cacagagtac ttattcattc atttcccaga tcatcatgaa
     5221 ggacacttaa ctttgttgcg aagctctttg gtgaataata gaactcaggc caaggtagcg
     5281 gaggagctgg gcatgcagga gtacgccata accaacgaca agaccaagag gcctgtggcg
     5341 cttcgcacca agaccttggc ggaccttttg gaatcattta ttgcagcgct gtacattgat
     5401 aaggatttgg aatatgttca tactttcatg aatgtctgct tctttccacg attgaaagag
     5461 ttcattttga atcaggattg gaatgacccc aaatcccagc ttcagcagtg ttgcttgaca
     5521 cttaggacag aaggaaaaga gccagacatt cctctgtaca agactctgca gacagtgggc
     5581 ccatcccatg cccgaaccta cactgtggct gtttatttca agggagaaag aataggctgt
     5641 gggaaaggac caagtattca gcaagcggaa atgggagcag caatggatgc gcttgaaaaa
     5701 tataattttc cccagatggc ccatcagaag cggttcatcg aacggaagta cagacaagag
     5761 ttaaaagaaa tgaggtggga aagagagcat caagagagag agccagatga gactgaagac
     5821 atcaagaaag actacaaaga cgatgacgat aaataatcta gagacaatca ctagtgaatt
     5881 cgcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     5941 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6001 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6061 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6121 atagcaggca tgctggggag tcgaaattca gaagaactcg tcaagaaggc gatagaaggc
     6181 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     6241 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     6301 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     6361 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     6421 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     6481 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     6541 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     6601 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     6661 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     6721 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     6781 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     6841 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     6901 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     6961 tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
     7021 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
     7081 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc
     7141 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     7201 catccggggt cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa
     7261 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7321 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7381 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7441 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7501 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7561 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7621 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7681 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7741 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7801 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7861 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7921 aggggggcgg agcctatgga aaaacgccag caacgcggc
//
