LOCUS       sequence-340158-        7567 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 7567)
  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 7567)
  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: p3xFLAG-CMV-Drosha (NLS-P426)
FEATURES             Location/Qualifiers
     source          1..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        355..558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     509..529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             588..653
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             657..677
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3565..4187
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3764..3783)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4211..4231)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4216..4545
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4396..4531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4583..4602)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4584..4602)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4616..4632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4616..4632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4629..4651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4640..4656
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4664..4670,4671..4688,4689..4694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4709..4730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4847..4864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5018..5606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5098..5117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5777..6568,6569..6637))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6400..6419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6638..6742)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6769..7224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6856..6875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7066..7087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7350..7372
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7364..7381
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7365..7381
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        join(7542..7567,1..354)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
       61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
      241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      301 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      361 cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      421 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      481 aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag
      541 gtctatataa gcagagctcg tttagtgaac cgtcagaatt aaccatggac tacaaagacc
      601 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagcttccaa
      661 agaagaaacg gaaggtcggt atcgccggaa agcttcccat ggatcaggtg ggagattcta
      721 cagtggttgg aacgagtagg cttcgtgact tatatgacaa atttgaggag gagttgggga
      781 gcaggcaaga aaaggccaaa gctgctcggc ctccgtggga acctccaaag acgaagctcg
      841 atgaagattt agagagttcc agtgaatccg agtgtgagtc tgatgaggac agcacctgtt
      901 ctagcagctc agactctgaa gtttttgacg ttattgcaga aatcaaacgc aaaaaggccc
      961 accctgaccg acttcatgat gaactttggt acaacgatcc aggccagatg aatgatggac
     1021 cactctgcaa atgcagcgca aaggcaagac gcacaggaat taggcacagc atttatcctg
     1081 gagaagaggc catcaagccc tgtcgtccta tgaccaacaa tgctggcaga cttttccact
     1141 accggatcac agtctccccg cctacgaact ttttaactga caggccaact gttatagaat
     1201 acgatgatca cgagtatatc tttgaaggat tttctatgtt tgcacatgcc cccctgacca
     1261 atattccact gtgtaaagta attagattca acatagacta cacgattcat ttcattgaag
     1321 agatgatgcc ggagaatttt tgtgtgaaag ggcttgaact cttttcactg ttcctattca
     1381 gagatatttt ggaattatat gactggaatc ttaaaggtcc tttgtttgaa gacagccctc
     1441 cctgctgccc aagatttcat ttcatgccac gttttgtaag atttcttcca gatggaggaa
     1501 aggaagtgct gtccatgcac cagattctcc tgtacttgtt aaggtgcagc aaagccctgg
     1561 tgcctgagga ggagattgcc aatatgcttc agtgggagga gctggagtgg cagaaatatg
     1621 cagaagaatg caaaggcatg attgttacca accctggggc gaaaccaagc tctgtccgta
     1681 tcgatcaact ggatcgtgaa cagttcaacc ccgatgtgat tacttttccg attatcgtcc
     1741 actttgggat acgccctgca cagttgagtt atgcaggaga cccacagtac caaaaactgt
     1801 ggaagagtta tgtgaaactt cgccacctcc tagcaaatag tcccaaagtc aaacaaactg
     1861 acaaacagaa gctggcacag agggaggaag ccctccaaaa aatacggcag aagaatacaa
     1921 tgagacgaga agtaacggtg gagctaagta gccaaggatt ctggaaaact ggcatccgtt
     1981 ctgatgtctg tcagcatgca atgatgctac ctgttctgac ccatcatatc cgctaccacc
     2041 aatgcctaat gcatttggac aagttgatag gatatacttt ccaagatcgt tgtctgttgc
     2101 agctggccat gactcatcca agtcatcatt taaattttgg aatgaatcct gatcatgcca
     2161 ggaattcatt atctaactgt ggaattcggc agcccaaata cggagacaga aaagttcatc
     2221 acatgcacat gcggaagaaa gggattaaca ccttgataaa tatcatgtca cgccttggcc
     2281 aagatgaccc aactccctcg aggattaacc acaatgaacg gttggaattc ctgggtgatg
     2341 ctgttgttga atttctgacc agcgtccatt tgtactattt gtttcctagt ctggaagaag
     2401 gaggattagc aacctatcgg actgccattg ttcagaatca gcaccttgcc atgctagcaa
     2461 agaaacttga actggatcga tttatgctgt atgctcacgg gcctgacctt tgtagagaat
     2521 cggaccttcg acatgcaatg gccaattgtt ttgaagcgtt aataggagct gtttacttgg
     2581 agggaagcct ggaggaagcc aagcagttat ttggacgctt gctctttaat gatccggacc
     2641 tgcgcgaagt ctggctcaat tatcctctcc acccactcca actacaagag ccaaatactg
     2701 atcgacaact tattgaaact tctccggttc tacaaaaact tactgagttt gaagaagcaa
     2761 ttggagtaat ttttactcat gttcgacttc tggcaagggc attcacattg agaactgtgg
     2821 gatttaacca tctgacccta ggccacaatc agagaatgga attcctaggt gactccataa
     2881 tgcaactggt agccacagag tacttattca ttcatttccc agatcatcat gaaggacact
     2941 taactttgtt gcgaagctct ttggtgaata atagaactca ggccaaggta gcggaggagc
     3001 tgggcatgca ggagtacgcc ataaccaacg acaagaccaa gaggcctgtg gcgcttcgca
     3061 ccaagacctt ggcggacctt ttggaatcat ttattgcagc gctgtacatt gataaggatt
     3121 tggaatatgt tcatactttc atgaatgtct gcttctttcc acgattgaaa gagttcattt
     3181 tgaatcagga ttggaatgac cccaaatccc agcttcagca gtgttgcttg acacttagga
     3241 cagaaggaaa agagccagac attcctctgt acaagactct gcagacagtg ggcccatccc
     3301 atgcccgaac ctacactgtg gctgtttatt tcaagggaga aagaataggc tgtgggaaag
     3361 gaccaagtat tcagcaagcg gaaatgggag cagcaatgga tgcgcttgaa aaatataatt
     3421 ttccccagat ggcccatcag aagcggttca tcgaacggaa gtacagacaa gagttaaaag
     3481 aaatgaggtg ggaaagagag catcaagaga gagagccaga tgagactgaa gacatcaaga
     3541 aataagtcga ctctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     3601 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3661 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3721 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3781 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3841 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3901 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3961 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     4021 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     4081 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     4141 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     4201 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4261 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4321 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4501 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa
     4561 ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat
     4621 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4681 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4741 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4801 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5701 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5761 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5821 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5881 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5941 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6001 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6061 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6121 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6181 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6241 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6301 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6361 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6421 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6481 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6541 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6601 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     6781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6841 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6961 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     7201 acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
     7261 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     7321 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     7381 tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta
     7441 tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat
     7501 gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat tattgactag
     7561 ttattaa
//
