LOCUS       sequence-340164-        7300 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 7300)
  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 7300)
  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(R871)
FEATURES             Location/Qualifiers
     source          1..7300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     67..89
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     81..98
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     82..98
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        259..638
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..842
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     793..813
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             872..937
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3582..4204
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3781..3800)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4228..4248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4233..4562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4413..4548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4600..4619)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4601..4619)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4633..4649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4633..4649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4646..4668)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4657..4673
                     /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(4681..4687,4688..4705,4706..4711))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4726..4747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4864..4881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5035..5623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5115..5134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5794..6585,6586..6654))
                     /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     6417..6436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6655..6759)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6786..7241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6873..6892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7083..7104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
       61 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc tatacattga
      121 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
      181 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
      241 caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      301 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      361 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      421 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      481 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
      541 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
      601 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      661 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      721 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc
      781 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      841 ctcgtttagt gaaccgtcag aattaaccat ggactacaaa gaccatgacg gtgattataa
      901 agatcatgac atcgattaca aggatgacga tgacaagctt gccgtggcga tgcagggaaa
      961 cacatgtcac agaatgtcgt tccgcccggg acgagggtgt ccccgaggac gaggaggaca
     1021 tggagccaga cccccagcac catcctttag gccccaaaat ctgaggctgc ttcaccctca
     1081 gcggcctcct gtgcaatatc aatatgaacc tccaagtgcc ccttccacca ctttctcaaa
     1141 ctctccagcc cccaattttc tccctccacg accagacttt gtacccttcc ccccacccat
     1201 gcctccgtca gcgcaaggcc ctcttccccc ctgcccaatc aggccgcctt tccccaacca
     1261 ccagatgagg caccccttcc cagttcctcc ttgttttcct cccatgccac caccaatgcc
     1321 ttgtcctaat aaccccccag tccctggggt acctcctgga caaggcactt tccccttcat
     1381 gatgccccct ccctccatgc ctcatccccc gccccctcca gtcatgccgc agcaggttaa
     1441 ttatcagtac cctccgggct attctcacca caacttccca cctcccagtt ttaatagttt
     1501 ccagaacaac cctagttctt tcctgcccag tgctaataac agcggtagtc ctcatttcag
     1561 acatctccct ccatacccac tcccaaaggc tcccagtgag agaaggtccc cagaaaggct
     1621 gaaacactat gatgaccaca ggcaccgaga tcacagtcat gggcgaggtg agaggcatcg
     1681 gtccctggat cggcgggagc gaggccgcag tcccgacagg agaagacaag acagccggta
     1741 cagatctgat tatgaccgag ggagaacacc atctcgccac cgcagctacg aacggagcag
     1801 agagcgagaa cgggagagac acaggcatcg agacaaccga agatcaccat ctctggaaag
     1861 gtcctacaaa aaagagtata agagatctgg aaggagttac ggtttattgg ttgttcctga
     1921 acctgctgga tgcacaccag aattacctgg ggagattatt aaaaatacag attcttgggc
     1981 cccacccctg gagattgtga atcatcgctc cccaagtagg gagaagaaga gagctcgttg
     2041 ggaggaagaa aaagaccgtt ggagtgacaa ccagagttct ggcaaagaca agaactatac
     2101 ctcaatcaag gaaaaagagc ccgaggagac catgcctgac aagaatgagg aggaagaaga
     2161 agaacttctt aagcctgtgt ggattcgatg cactcattca gaaaactact actccagtga
     2221 ccccatggat caggtgggag attctacagt ggttggaacg agtaggcttc gtgatttata
     2281 tgacaaattt gaggaggagt tggggagcag gcaagaaaag gccaaagctg ctcggcctcc
     2341 gtgggaacct ccaaagacga agctcgatga agatttagag agttccagtg aatccgagtg
     2401 tgagtctgat gaggacagca cctgttctag cagctcagac tctgaagttt ttgacgttat
     2461 tgcagaaatc aaacgcaaaa aggcccaccc tgaccgactt catgatgaac tttggtacaa
     2521 cgatccaggc cagatgaatg atggaccact ctgcaaatgc agcgcaaagg caagacgcac
     2581 aggaattagg cacagcattt atcctggaga agaggccatc aagccctgtc gtcctatgac
     2641 caacaatgct ggcagacttt tccactaccg gatcacagtc tccccgccta cgaacttttt
     2701 aactgacagg ccaactgtta tagaatacga tgatcacgag tatatctttg agggattttc
     2761 tatgtttgca catgcccccc tgaccaatat tccactgtgt aaagtaatta gattcaacat
     2821 agactacacg attcatttca ttgaagagat gatgccggag aatttttgtg tgaaagggct
     2881 tgaactcttt tcactgttcc tattcagaga tattttggaa ttatatgact ggaatcttaa
     2941 aggtcctttg tttgaagaca gccctccctg ctgcccaaga tttcatttca tgccacgttt
     3001 tgtaagattt cttccagatg gaggaaagga agtgctgtcc atgcaccaga ttctcctgta
     3061 cttgttaagg tgcagcaaag ccctggtgcc tgaggaggag attgccaata tgcttcagtg
     3121 ggaggagctg gagtggcaga aatatgcaga agaatgcaaa ggcatgattg ttaccaaccc
     3181 tgggacgaaa ccaagctctg tccgtatcga tcaactggat cgtgaacagt tcaaccccga
     3241 tgtgattact tttccgatta tcgtccactt tgggatacgc cctgcacagt tgagttatgc
     3301 aggagaccca cagtaccaaa aactgtggaa gagttatgtg aaacttcgcc acctcctagc
     3361 aaatagtccc aaagtcaaac aaactgacaa acagaagctg gcacagaggg aggaagccct
     3421 ccaaaaaata cggcagaaga atacaatgag acgagaagta acggtggagc taagtagcca
     3481 aggattctgg aaaactggca tccgttctga tgtctgtcag catgcaatga tgctacctgt
     3541 tctgacccat catatccgct aagtcgactc tagaggatcc cgggtggcat ccctgtgacc
     3601 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3661 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3721 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     3781 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3841 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3901 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3961 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     4021 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     4081 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     4141 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     4201 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4321 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4381 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4441 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4501 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4561 aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa
     4621 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4681 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4741 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4801 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4861 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4921 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4981 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5041 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5101 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5161 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5221 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5281 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5341 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5401 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5461 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5521 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5581 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5641 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5701 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5761 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5821 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5881 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5941 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6001 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6061 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6121 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6181 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6241 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6301 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6361 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6421 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6481 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6541 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6601 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6661 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6721 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6781 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6841 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6901 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6961 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     7021 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     7081 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     7141 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     7201 atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc attcaggctg
     7261 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
//
