LOCUS       sequence-340150-        8809 bp    DNA     circular SYN 15-APR-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 Apr 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(W1258)
FEATURES             Location/Qualifiers
     source          1..8809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             766..831
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4985..5607
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5184..5203)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5631..5651)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5636..5965
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5816..5951
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6003..6022)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6004..6022)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6036..6052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6036..6052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6049..6071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6060..6076
                     /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(6084..6090,6091..6108,6109..6114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6129..6150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6267..6284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6438..7026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6518..6537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7197..7988,7989..8057))
                     /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     7820..7839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8058..8162)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8189..8644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8276..8295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8486..8507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8770..8792
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8784..8801
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8785..8801
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
       61 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
      121 tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctcgtt tagtgaaccg tcagaattaa ccatggacta caaagaccat
      781 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gcttgccatg
      841 gcgatgcagg gaaacacatg tcacagaatg tcgttccacc cgggacgagg gtgtccccga
      901 ggacgaggag gacatggagc cagaccccca gcaccatcct ttaggcccca aaatctgagg
      961 ctgcttcacc ctcagcagcc tcctgtgcaa tatcaatatg aacctccaag tgccccttcc
     1021 accactttct caaactctcc agcccccaat tttctccctc cacgaccaga ctttgtaccc
     1081 ttccccccac ccatgcctcc gtcagcgcaa ggccctcttc ccccctgccc aatcaggccg
     1141 cctttcccca accaccagat gaggcacccc ttcccagttc ctccttgttt tcctcccatg
     1201 ccaccaccaa tgccttgtcc taataacccc ccagtccctg gggtacctcc tggacaaggc
     1261 actttcccct tcatgatgcc ccctccctcc atgcctcatc ccccgccccc tccagtcatg
     1321 ccgcagcagg ttaattatca gtaccctccg ggctattctc accacaactt cccacctccc
     1381 agttttaata gtttccagaa caaccctagt tctttcctgc ccagtgctaa taacagcggt
     1441 agtcctcatt tcagacatct ccctccatac ccactcccaa aggctcccag tgagagaagg
     1501 tccccagaaa ggctgaaaca ctatgatgac cacaggcacc gagatcacag tcatgggcga
     1561 ggtgagaggc atcggtccct ggatcggcgg gagcgaggcc gcagtcccga caggagaaga
     1621 caagacagcc ggtacagatc tgattatgac cgagggagaa caccatctcg ccaccgcagc
     1681 tacgaacgga gcagagagcg agaacgggag agacacaggc atcgagacaa ccgaagatca
     1741 ccatctctgg aaaggtccta caaaaaagag tataagagat ctggaaggag ttacggttta
     1801 ttggttgttc ctgaacctgc tggatgcaca ccagaattac ctggggagat tattaaaaat
     1861 acagattctt gggccccacc cctggagatt gtgaatcatc gctccccaag tagggagaag
     1921 aagagagctc gttgggagga agaaaaagac cgttggagtg acaaccagag ttctggcaaa
     1981 gacaagaact atacctcaat caaggaaaaa gagcccgagg agaccatgcc tgacaagaat
     2041 gaggaggaag aagaagaact tcttaagcct gtgtggattc gatgcactca ttcagaaaac
     2101 tactactcca gtgaccccat ggatcaggtg ggagattcta cagtggttgg aacgagtagg
     2161 cttcgtgact tatatgacaa atttgaggag gagttgggga gcaggcaaga aaaggccaaa
     2221 gctgctcggc ctccgtggga acctccaaag acgaagctcg atgaagattt agagagttcc
     2281 agtgaatccg agtgtgagtc tgatgaggac agcacctgtt ctagcagctc agactctgaa
     2341 gtttttgacg ttattgcaga aatcaaacgc aaaaaggccc accctgaccg acttcatgat
     2401 gaactttggt acaacgatcc aggccagatg aatgatggac cactctgcaa atgcagcgca
     2461 aaggcaagac gcacaggaat taggcacagc atttatcctg gagaagaggc catcaagccc
     2521 tgtcgtccta tgaccaacaa tgctggcaga cttttccact accggatcac agtctccccg
     2581 cctacgaact ttttaactga caggccaact gttatagaat acgatgatca cgagtatatc
     2641 tttgaaggat tttctatgtt tgcacatgcc cccctgacca atattccact gtgtaaagta
     2701 attagattca acatagacta cacgattcat ttcattgaag agatgatgcc ggagaatttt
     2761 tgtgtgaaag ggcttgaact cttttcactg ttcctattca gagatatttt ggaattatat
     2821 gactggaatc ttaaaggtcc tttgtttgaa gacagccctc cctgctgccc aagatttcat
     2881 ttcatgccac gttttgtaag atttcttcca gatggaggaa aggaagtgct gtccatgcac
     2941 cagattctcc tgtacttgtt aaggtgcagc aaagccctgg tgcctgagga ggagattgcc
     3001 aatatgcttc agtgggagga gctggagtgg cagaaatatg cagaagaatg caaaggcatg
     3061 attgttacca accctgggac gaaaccaagc tctgtccgta tcgatcaact ggatcgtgaa
     3121 cagttcaacc ccgatgtgat tacttttccg attatcgtcc actttgggat acgccctgca
     3181 cagttgagtt atgcaggaga cccacagtac caaaaactgt ggaagagtta tgtgaaactt
     3241 cgccacctcc tagcaaatag tcccaaagtc aaacaaactg acaaacagaa gctggcacag
     3301 agggaggaag ccctccaaaa aatacggcag aagaatacaa tgagacgaga agtaacggtg
     3361 gagctaagta gccaaggatt ctggaaaact ggcatccgtt ctgatgtctg tcagcatgca
     3421 atgatgctac ctgttctgac ccatcatatc cgctaccacc aatgcctaat gcatttggac
     3481 aagttgatag gatatacttt ccaagatcgt tgtctgttgc agctggccat gactcatcca
     3541 agtcatcatt taaattttgg aatgaatcct gatcatgcca ggaattcatt atctaactgt
     3601 ggaattcggc agcccaaata cggagacaga aaagttcatc acatgcacat gcggaagaaa
     3661 gggattaaca ccttgataaa tatcatgtca cgccttggcc aagatgaccc aactccctcg
     3721 aggattaacc acaatgaacg gttggaattc ctgggtgatg ctgttgttga atttctgacc
     3781 agcgtccatt tgtactattt gtttcctagt ctggaagaag gaggattagc aacctatcgg
     3841 actgccattg ttcagaatca gcaccttgcc atgctagcaa agaaacttga actggatcga
     3901 tttatgctgt atgctcacgg gcctgacctt tgtagagaat cggaccttcg acatgcaatg
     3961 gccaattgtt ttgaagcgtt aataggagct gtttacttgg agggaagcct ggaggaagcc
     4021 aagcagttat ttggacgctt gctctttaat gatccggacc tgcgcgaagt ctggctcaat
     4081 tatcctctcc acccactcca actacaagag ccaaatactg atcgacaact tattgaaact
     4141 tctccggttc tacaaaaact tactgagttt gaagaagcaa ttggagtaat ttttactcat
     4201 gttcgacttc tggcaagggc attcacattg agaactgtgg gatttaacca tctgacccta
     4261 ggccacaatc agagaatgga attcctaggt gactccataa tgcaactggt agccacagag
     4321 tacttattca ttcatttccc agatcatcat gaaggacact taactttgtt gcgaagctct
     4381 ttggtgaata atagaactca ggccaaggta gcggaggagc tgggcatgca ggagtacgcc
     4441 ataaccaacg acaagaccaa gaggcctgtg gcgcttcgca ccaagacctt ggcggacctt
     4501 ttggaatcat ttattgcagc gctgtacatt gataaggatt tggaatatgt tcatactttc
     4561 atgaatgtct gcttctttcc acgattgaaa gagttcattt tgaatcagga ttagaatgac
     4621 cccaaatccc agcttcagca gtgttgcttg acacttagga cagaaggaaa agagccagac
     4681 attcctctgt acaagactct gcagacagtg ggcccatccc atgcccgaac ctacactgtg
     4741 gctgtttatt tcaagggaga aagaataggc tgtgggaaag gaccaagtat tcagcaagcg
     4801 gaaatgggag cagcaatgga tgcgcttgaa aaatataatt ttccccagat ggcccatcag
     4861 aagcggttca tcgaacggaa gtacagacaa gagttaaaag aaatgaggtg ggaaagagag
     4921 catcaagaga gagagccaga tgagactgaa gacatcaaga aataagtcga ctctagagga
     4981 tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     5041 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     5101 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     5161 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     5221 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     5281 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     5341 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     5401 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     5461 tctgatttta aaataactat accagcagga ggacgtccag acacagcata ggctacctgg
     5521 ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg
     5581 gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg
     5641 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5701 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5761 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5821 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5881 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5941 ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa
     6001 ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     6061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8161 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     8221 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     8281 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8341 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8401 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8461 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     8521 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     8581 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     8641 atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     8701 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     8761 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagct
//
