LOCUS       sequence-339754-        8809 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 8809)
  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 8809)
  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 (E419)
FEATURES             Location/Qualifiers
     source          1..8809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..115
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     107..124
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     108..124
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        285..664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        665..868
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     819..839
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             898..963
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    5117..5739
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5316..5335)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5763..5783)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5768..6097
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5948..6083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6135..6154)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6136..6154)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6168..6184)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6168..6184)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6181..6203)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6192..6208
                     /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(6216..6222,6223..6240,6241..6246))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6261..6282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6399..6416)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6570..7158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6650..6669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7329..8120,8121..8189))
                     /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     7952..7971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8190..8294)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8321..8776
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8408..8427)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8618..8639
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
       61 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      121 cagtgccaag ctgatctata cattgaatca atattggcaa ttagccatat tagtcattgg
      181 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
      241 tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac
      301 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      361 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      541 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      601 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      661 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      721 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      781 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
      841 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagaatt aaccatggac
      901 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
      961 aagcttgcca tggcgatgca gggaaacaca tgtcacagaa tgtcgttcca cccgggacga
     1021 gggtgtcccc gaggacgagg aggacatgga gccagacccc cagcaccatc ctttaggccc
     1081 caaaatctga ggctgcttca ccctcagcag cctcctgtgc aatatcaata tgaacctcca
     1141 agtgcccctt ccaccacttt ctcaaactct ccagccccca attttctccc tccacgacca
     1201 gactttgtac ccttcccccc acccatgcct ccgtcagcgc aaggccctct tcccccctgc
     1261 ccaatcaggc cgcctttccc caaccaccag atgaggcacc ccttcccagt tcctccttgt
     1321 tttcctccca tgccaccacc aatgccttgt cctaataacc ccccagtccc tggggtacct
     1381 cctggacaag gcactttccc cttcatgatg ccccctccct ccatgcctca tcccccgccc
     1441 cctccagtca tgccgcagca ggttaattat cagtaccctc cgggctattc tcaccacaac
     1501 ttcccacctc ccagttttaa tagtttccag aacaacccta gttctttcct gcccagtgct
     1561 aataacagcg gtagtcctca tttcagacat ctccctccat acccactccc aaaggctccc
     1621 agtgagagaa ggtccccaga aaggctgaaa cactatgatg accacaggca ccgagatcac
     1681 agtcatgggc gaggtgagag gcatcggtcc ctggatcggc gggagcgagg ccgcagtccc
     1741 gacaggagaa gacaagacag ccggtacaga tctgattatg accgagggag aacaccatct
     1801 cgccaccgca gctacgaacg gagcagagag cgagaacggg agagacacag gcatcgagac
     1861 aaccgaagat caccatctct ggaaaggtcc tacaaaaaag agtataagag atctggaagg
     1921 agttacggtt tattggttgt tcctgaacct gctggatgca caccagaatt acctggggag
     1981 attattaaaa atacagattc ttgggcccca cccctggaga ttgtgaatca tcgctcccca
     2041 agtagggaga agaagagagc tcgttgggag gaagaaaaag accgttggag tgacaaccag
     2101 agttctggca aagacaagaa ctatacctca atcaaggaaa aagagcccga ggagaccatg
     2161 cctgacaaga atgaggagga agaagaagaa cttcttaagc ctgtgtggat tcgatgcact
     2221 cattcataaa actactactc cagtgacccc atggatcagg tgggagattc tacagtggtt
     2281 ggaacgagta ggcttcgtga cttatatgac aaatttgagg aggagttggg gagcaggcaa
     2341 gaaaaggcca aagctgctcg gcctccgtgg gaacctccaa agacgaagct cgatgaagat
     2401 ttagagagtt ccagtgaatc cgagtgtgag tctgatgagg acagcacctg ttctagcagc
     2461 tcagactctg aagtttttga cgttattgca gaaatcaaac gcaaaaaggc ccaccctgac
     2521 cgacttcatg atgaactttg gtacaacgat ccaggccaga tgaatgatgg accactctgc
     2581 aaatgcagcg caaaggcaag acgcacagga attaggcaca gcatttatcc tggagaagag
     2641 gccatcaagc cctgtcgtcc tatgaccaac aatgctggca gacttttcca ctaccggatc
     2701 acagtctccc cgcctacgaa ctttttaact gacaggccaa ctgttataga atacgatgat
     2761 cacgagtata tctttgaagg attttctatg tttgcacatg cccccctgac caatattcca
     2821 ctgtgtaaag taattagatt caacatagac tacacgattc atttcattga agagatgatg
     2881 ccggagaatt tttgtgtgaa agggcttgaa ctcttttcac tgttcctatt cagagatatt
     2941 ttggaattat atgactggaa tcttaaaggt cctttgtttg aagacagccc tccctgctgc
     3001 ccaagatttc atttcatgcc acgttttgta agatttcttc cagatggagg aaaggaagtg
     3061 ctgtccatgc accagattct cctgtacttg ttaaggtgca gcaaagccct ggtgcctgag
     3121 gaggagattg ccaatatgct tcagtgggag gagctggagt ggcagaaata tgcagaagaa
     3181 tgcaaaggca tgattgttac caaccctggg acgaaaccaa gctctgtccg tatcgatcaa
     3241 ctggatcgtg aacagttcaa ccccgatgtg attacttttc cgattatcgt ccactttggg
     3301 atacgccctg cacagttgag ttatgcagga gacccacagt accaaaaact gtggaagagt
     3361 tatgtgaaac ttcgccacct cctagcaaat agtcccaaag tcaaacaaac tgacaaacag
     3421 aagctggcac agagggagga agccctccaa aaaatacggc agaagaatac aatgagacga
     3481 gaagtaacgg tggagctaag tagccaagga ttctggaaaa ctggcatccg ttctgatgtc
     3541 tgtcagcatg caatgatgct acctgttctg acccatcata tccgctacca ccaatgccta
     3601 atgcatttgg acaagttgat aggatatact ttccaagatc gttgtctgtt gcagctggcc
     3661 atgactcatc caagtcatca tttaaatttt ggaatgaatc ctgatcatgc caggaattca
     3721 ttatctaact gtggaattcg gcagcccaaa tacggagaca gaaaagttca tcacatgcac
     3781 atgcggaaga aagggattaa caccttgata aatatcatgt cacgccttgg ccaagatgac
     3841 ccaactccct cgaggattaa ccacaatgaa cggttggaat tcctgggtga tgctgttgtt
     3901 gaatttctga ccagcgtcca tttgtactat ttgtttccta gtctggaaga aggaggatta
     3961 gcaacctatc ggactgccat tgttcagaat cagcaccttg ccatgctagc aaagaaactt
     4021 gaactggatc gatttatgct gtatgctcac gggcctgacc tttgtagaga atcggacctt
     4081 cgacatgcaa tggccaattg ttttgaagcg ttaataggag ctgtttactt ggagggaagc
     4141 ctggaggaag ccaagcagtt atttggacgc ttgctcttta atgatccgga cctgcgcgaa
     4201 gtctggctca attatcctct ccacccactc caactacaag agccaaatac tgatcgacaa
     4261 cttattgaaa cttctccggt tctacaaaaa cttactgagt ttgaagaagc aattggagta
     4321 atttttactc atgttcgact tctggcaagg gcattcacat tgagaactgt gggatttaac
     4381 catctgaccc taggccacaa tcagagaatg gaattcctag gtgactccat aatgcaactg
     4441 gtagccacag agtacttatt cattcatttc ccagatcatc atgaaggaca cttaactttg
     4501 ttgcgaagct ctttggtgaa taatagaact caggccaagg tagcggagga gctgggcatg
     4561 caggagtacg ccataaccaa cgacaagacc aagaggcctg tggcgcttcg caccaagacc
     4621 ttggcggacc ttttggaatc atttattgca gcgctgtaca ttgataagga tttggaatat
     4681 gttcatactt tcatgaatgt ctgcttcttt ccacgattga aagagttcat tttgaatcag
     4741 gattggaatg accccaaatc ccagcttcag cagtgttgct tgacacttag gacagaagga
     4801 aaagagccag acattcctct gtacaagact ctgcagacag tgggcccatc ccatgcccga
     4861 acctacactg tggctgttta tttcaaggga gaaagaatag gctgtgggaa aggaccaagt
     4921 attcagcaag cggaaatggg agcagcaatg gatgcgcttg aaaaatataa ttttccccag
     4981 atggcccatc agaagcggtt catcgaacgg aagtacagac aagagttaaa agaaatgagg
     5041 tgggaaagag agcatcaaga gagagagcca gatgagactg aagacatcaa gaaataagtc
     5101 gactctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     5161 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     5221 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     5281 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     5341 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     5401 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     5461 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     5521 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     5581 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     5641 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     5701 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     5761 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5821 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5881 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5941 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6001 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6061 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa
     6121 ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt
     6181 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6241 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6301 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6361 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6421 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6481 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6541 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6601 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6661 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6721 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6781 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6841 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6901 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6961 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7021 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7081 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7141 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7201 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7261 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7321 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7381 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7441 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7501 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7561 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7621 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7681 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7741 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7801 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7861 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7921 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7981 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8041 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8101 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8161 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8221 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8281 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     8341 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     8401 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     8461 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     8521 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     8581 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     8641 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     8701 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     8761 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttggga
//
