LOCUS       sequence-340155-        7528 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 7528)
  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 7528)
  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 (P426)
FEATURES             Location/Qualifiers
     source          1..7528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        168..547
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        548..751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     702..722
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             781..846
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3719..4341
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3918..3937)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4365..4385)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4370..4699
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4550..4685
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4737..4756)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4738..4756)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4770..4786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4770..4786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4783..4805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4794..4810
                     /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(4818..4824,4825..4842,4843..4848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4863..4884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5001..5018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5172..5760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5252..5271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5931..6722,6723..6791))
                     /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     6554..6573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6792..6896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6923..7378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7010..7029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7220..7241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7504..7526
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7518..7528,1..7)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(7519..7528,1..7)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat
       61 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
      121 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt
      181 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      241 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      301 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      361 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      421 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      481 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      541 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
      601 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      661 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
      721 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga attaaccatg
      781 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
      841 gacaagcttc ccatggatca ggtgggagat tctacagtgg ttggaacgag taggcttcgt
      901 gacttatatg acaaatttga ggaggagttg gggagcaggc aagaaaaggc caaagctgct
      961 cggcctccgt gggaacctcc aaagacgaag ctcgatgaag atttagagag ttccagtgaa
     1021 tccgagtgtg agtctgatga ggacagcacc tgttctagca gctcagactc tgaagttttt
     1081 gacgttattg cagaaatcaa acgcaaaaag gcccaccctg accgacttca tgatgaactt
     1141 tggtacaacg atccaggcca gatgaatgat ggaccactct gcaaatgcag cgcaaaggca
     1201 agacgcacag gaattaggca cagcatttat cctggagaag aggccatcaa gccctgtcgt
     1261 cctatgacca acaatgctgg cagacttttc cactaccgga tcacagtctc cccgcctacg
     1321 aactttttaa ctgacaggcc aactgttata gaatacgatg atcacgagta tatctttgaa
     1381 ggattttcta tgtttgcaca tgcccccctg accaatattc cactgtgtaa agtaattaga
     1441 ttcaacatag actacacgat tcatttcatt gaagagatga tgccggagaa tttttgtgtg
     1501 aaagggcttg aactcttttc actgttccta ttcagagata ttttggaatt atatgactgg
     1561 aatcttaaag gtcctttgtt tgaagacagc cctccctgct gcccaagatt tcatttcatg
     1621 ccacgttttg taagatttct tccagatgga ggaaaggaag tgctgtccat gcaccagatt
     1681 ctcctgtact tgttaaggtg cagcaaagcc ctggtgcctg aggaggagat tgccaatatg
     1741 cttcagtggg aggagctgga gtggcagaaa tatgcagaag aatgcaaagg catgattgtt
     1801 accaaccctg gggcgaaacc aagctctgtc cgtatcgatc aactggatcg tgaacagttc
     1861 aaccccgatg tgattacttt tccgattatc gtccactttg ggatacgccc tgcacagttg
     1921 agttatgcag gagacccaca gtaccaaaaa ctgtggaaga gttatgtgaa acttcgccac
     1981 ctcctagcaa atagtcccaa agtcaaacaa actgacaaac agaagctggc acagagggag
     2041 gaagccctcc aaaaaatacg gcagaagaat acaatgagac gagaagtaac ggtggagcta
     2101 agtagccaag gattctggaa aactggcatc cgttctgatg tctgtcagca tgcaatgatg
     2161 ctacctgttc tgacccatca tatccgctac caccaatgcc taatgcattt ggacaagttg
     2221 ataggatata ctttccaaga tcgttgtctg ttgcagctgg ccatgactca tccaagtcat
     2281 catttaaatt ttggaatgaa tcctgatcat gccaggaatt cattatctaa ctgtggaatt
     2341 cggcagccca aatacggaga cagaaaagtt catcacatgc acatgcggaa gaaagggatt
     2401 aacaccttga taaatatcat gtcacgcctt ggccaagatg acccaactcc ctcgaggatt
     2461 aaccacaatg aacggttgga attcctgggt gatgctgttg ttgaatttct gaccagcgtc
     2521 catttgtact atttgtttcc tagtctggaa gaaggaggat tagcaaccta tcggactgcc
     2581 attgttcaga atcagcacct tgccatgcta gcaaagaaac ttgaactgga tcgatttatg
     2641 ctgtatgctc acgggcctga cctttgtaga gaatcggacc ttcgacatgc aatggccaat
     2701 tgttttgaag cgttaatagg agctgtttac ttggagggaa gcctggagga agccaagcag
     2761 ttatttggac gcttgctctt taatgatccg gacctgcgcg aagtctggct caattatcct
     2821 ctccacccac tccaactaca agagccaaat actgatcgac aacttattga aacttctccg
     2881 gttctacaaa aacttactga gtttgaagaa gcaattggag taatttttac tcatgttcga
     2941 cttctggcaa gggcattcac attgagaact gtgggattta accatctgac cctaggccac
     3001 aatcagagaa tggaattcct aggtgactcc ataatgcaac tggtagccac agagtactta
     3061 ttcattcatt tcccagatca tcatgaagga cacttaactt tgttgcgaag ctctttggtg
     3121 aataatagaa ctcaggccaa ggtagcggag gagctgggca tgcaggagta cgccataacc
     3181 aacgacaaga ccaagaggcc tgtggcgctt cgcaccaaga ccttggcgga ccttttggaa
     3241 tcatttattg cagcgctgta cattgataag gatttggaat atgttcatac tttcatgaat
     3301 gtctgcttct ttccacgatt gaaagagttc attttgaatc aggattggaa tgaccccaaa
     3361 tcccagcttc agcagtgttg cttgacactt aggacagaag gaaaagagcc agacattcct
     3421 ctgtacaaga ctctgcagac agtgggccca tcccatgccc gaacctacac tgtggctgtt
     3481 tatttcaagg gagaaagaat aggctgtggg aaaggaccaa gtattcagca agcggaaatg
     3541 ggagcagcaa tggatgcgct tgaaaaatat aattttcccc agatggccca tcagaagcgg
     3601 ttcatcgaac ggaagtacag acaagagtta aaagaaatga ggtgggaaag agagcatcaa
     3661 gagagagagc cagatgagac tgaagacatc aagaaataag tcgactctag aggatcccgg
     3721 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     3781 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     3841 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     3901 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     3961 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     4021 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     4081 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     4141 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     4201 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     4261 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     4321 tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt
     4381 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4441 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4501 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4561 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4621 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4681 aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag ttaattccct
     4741 atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     4801 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4921 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4981 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5581 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5881 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5941 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6001 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6061 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6121 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6181 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6241 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6301 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6361 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6421 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6481 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6541 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6601 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6661 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6721 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6781 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6841 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6901 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6961 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     7021 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     7081 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     7141 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     7201 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     7261 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     7321 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     7381 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     7441 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     7501 tttcccagtc acgacgttgt aaaacgac
//
