LOCUS       sequence-201136-        8841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  wild-type human Drosha cDNA is subcloned into pEGFP-C1 vector 
            between HindIII and KpnI sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8841)
  AUTHORS   Tang X, Zhang Y, Tucker L, Ramratnam B
  TITLE     Phosphorylation of the RNase III enzyme Drosha at Serine300 or 
            Serine302 is required for its nuclear localization.
  JOURNAL   Nucleic Acids Res. 2010 Oct;38(19):6610-9. doi: 10.1093/nar/gkq547. 
            Epub 2010 Jun 16.
  PUBMED    20554852
REFERENCE   2  (bases 1 to 8841)
  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: 
            "10.1093/nar/gkq547"; journalName: "Nucleic Acids Res"; date: 
            "2010-10"; volume: "38"; issue: "19"; pages: "6610-9"
COMMENT     Sequence Label: GFP-Drosha
FEATURES             Location/Qualifiers
     source          1..8841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5569..5690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5606..5625)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5660..5679
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5697..6152)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5834..5855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6046..6065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6179..6283
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6285..6642
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6285..6476
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6308..6328)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6493..6628
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6677..7471
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6677..7471
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6731..6750)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7341..7360
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7659..7678)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7703..7750
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8079..8667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8568..8587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga tgatgcaggg aaacacatgt
     1321 cacagaatgt cgttccaccc gggacgaggg tgtccccgag gacgaggagg acatggagcc
     1381 agaccctcag caccatcctt taggccccaa aatctgaggc tgcttcaccc tcagcagcct
     1441 cctgtgcaat atcaatatga acctccaagt gccccttcca ccactttctc aaactctcca
     1501 gcccccaatt ttctccctcc acgaccagac tttgtaccct tccccccacc catgcctccg
     1561 tcagcgcaag gccctcttcc cccctgccca atcaggccgc ctttccccaa ccaccagatg
     1621 aggcacccct tcccagttcc tccttgtttt cctcccatgc caccaccaat gccttgtcct
     1681 aataaccccc cagtccctgg ggcacctcct ggacaaggca ctttcccctt catgatgccc
     1741 cctccctcca tgcctcatcc cccgccccct ccagtcatgc cgcagcaggt taattatcag
     1801 taccctccgg gctattctca ccacaacttc ccacctccca gttttaatag tttccagaac
     1861 aaccctagtt ctttcctgcc cagtgctaat aacagcagta gtcctcattt cagacatctc
     1921 cctccatacc cactcccaaa ggctcccagt gagagaaggt ccccagaaag gctgaaacac
     1981 tatgatgacc acaggcaccg agatcacagt catgggcgag gtgagaggca tcggtccctg
     2041 gatcggcggg agcgaggccg cagtcccgac aggagaagac aagacagccg gtacagatct
     2101 gattatgacc gagggagaac accatctcgc cgccgcagct acgaacggag cagagagcga
     2161 gaacgggaga gacacaggca tcgagacaac cgaagatcac catctctgga aaggtcctac
     2221 aaaaaagagt ataagagatc tggaaggagt tacggtttat cggttgttcc tgaacctgct
     2281 ggatgcacac cagaattacc tggggagatt attaaaaata cagattcttg ggccccaccc
     2341 ctggagattg tgaatcatcg ctccccaagt agggagaaga agagagctcg ttgggaggaa
     2401 gaaaaagacc gttggagtga caaccagagt tctggcaaag acaagaacta tacctcaatc
     2461 aaggaaaaag agcccgagga gaccatgcct gacaagaatg aggaggaaga agaagaactt
     2521 cttaagcctg tgtggattcg atgcactcat tcagaaaact actactccag tgaccccatg
     2581 gatcaggtgg gagattctac agtggttgga acgagtaggc ttcgtgactt atatgacaaa
     2641 tttgaggagg agttggggag caggcaagaa aaggccaaag ctgctcggcc tccgtgggaa
     2701 cctccaaaga cgaagctcga tgaagattta gagagttcca gtgaatccga gtgtgagtct
     2761 gatgaggaca gcacctgttc tagcagctca gactctgaag tttttgacgt tattgcagaa
     2821 atcaaacgca aaaaggccca ccctgaccga cttcatgatg aactttggta caacgatcca
     2881 ggccagatga atgatggacc actctgcaaa tgcagcgcaa aggcaagacg cacaggaatt
     2941 aggcacagca tttatcctgg agaagaggcc atcaagccct gtcgtcctat gaccaacaat
     3001 gctggcagac ttttccacta ccggatcaca gtctccccgc ctacgaactt tttaactgac
     3061 aggccaactg ttatagaata cgatgatcac gagtatatct ttgaaggatt ttctatgttt
     3121 gcacatgccc ccctgaccaa tattccactg tgtaaagtaa ttagattcaa catagactac
     3181 acgattcatt tcattgaaga gatgatgccg gagaattttt gtgtgaaagg gcttgaactc
     3241 ttttcactgt tcctattcag agatattttg gaattatatg actggaatct taaaggtcct
     3301 ttgtttgaag acagccctcc ctgctgccca agatttcatt tcatgccacg ttttgtaaga
     3361 tttcttccag atggaggaaa ggaagtgctg tccatgcacc agattctcct gtacttgtta
     3421 aggtgcagca aagccctggt gcctgaggag gagattgcca atatgcttca gtgggaggag
     3481 ctggagtggc agaaatatgc agaagaatgc aaaggcatga ttgttaccaa ccctgggacg
     3541 aaaccaagct ctgtccgtat cgatcaactg gatcgtgaac agttcaaccc cgatgtgatt
     3601 acttttccga ttatcgtcca ctttgggata cgccctgcac agttgagtta tgcaggagac
     3661 ccacagtacc aaaaactgtg gaagagttat gtgaaacttc gccacctcct agcaaatagt
     3721 cccaaagtca aacaaactga caaacagaag ctggcacaga gggaggaagc cctccaaaaa
     3781 atacggcaga agaatacaat gagacgagaa gtaacggtgg agctaagtag ccaaggattc
     3841 tggaaaactg gcatccgttc tgatgtctgt cagcatgcaa tgatgctacc tgttctgacc
     3901 catcatatcc gctaccacca atgcctaatg catttggaca agttgatagg atatactttc
     3961 caagatcgtt gtctgttgca gctggccatg actcatccaa gtcatcattt aaattttgga
     4021 atgaatcctg atcatgccag gaattcatta tctaactgtg gaattcggca gcccaaatac
     4081 ggagacagaa aagttcatca catgcacatg cggaagaaag ggattaacac cttgataaat
     4141 atcatgtcac gccttggcca agatgaccca actccctcga ggattaacca caatgaacgg
     4201 ttggaattcc tgggtgatgc tgttgttgaa tttctgacca gcgtccattt gtactatttg
     4261 tttcctagtc tggaagaagg aggattagca acctatcgga ctgccattgt tcagaatcag
     4321 caccttgcca tgctagcaaa gaaacttgaa ctggatcgat ttatgctgta tgctcacggg
     4381 cctgaccttt gtagagaatc ggaccttcga catgcaatgg ccaattgttt tgaagcgtta
     4441 ataggagctg tttacttgga gggaagcctg gaggaagcca agcagttatt tggacgcttg
     4501 ctctttaatg atccggacct gcgcgaagtc tggctcaatt atcctctcca cccactccaa
     4561 ctacaagagc caaatactga tcgacaactt attgaaactt ctccggttct acaaaaactt
     4621 actgagtttg aagaagcaat tggagtaatt tttactcatg ttcgacttct ggcaagggca
     4681 ttcacattga gaactgtggg atttaaccat ctgaccctag gccacaatca gagaatggaa
     4741 ttcctaggtg actccataat gcaactggta gccacagagt acttattcat tcatttccca
     4801 gatcatcatg aaggacactt aactttgttg cgaagctctt tggtgaataa tagaactcag
     4861 gccaaggtag cggaggagct gggcatgcag gagtacgcca taaccaacga caagaccaag
     4921 aggcctgtgg cgcttcgcac caagaccttg gcggaccttt tggaatcatt tattgcagcg
     4981 ctgtacattg ataaggattt ggaatatgtt catactttca tgaatgtctg cttctttcca
     5041 cgattgaaag agttcatttt gaatcaggat tggaatgacc ccaaatccca gcttcagcag
     5101 tgttgcttga cacttaggac agaaggaaaa gagccagaca ttcctctgta caagactctg
     5161 cagacagtgg gcccatccca tgcccgaacc tacactgtgg ctgtttattt caagggagaa
     5221 agaataggct gtgggaaagg accaagtatt cagcaagcgg aaatgggagc agcaatggat
     5281 gcgcttgaaa aatataattt tccccagatg gcccatcaga agcggttcat cgaacggaag
     5341 tacagacaag agttaaaaga aatgaggtgg gaaagagagc atcaagagag agagccagat
     5401 gagactgaag acatcaagaa ataaggtacc gcgggcccgg gatccaccgg atctagataa
     5461 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     5521 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     5581 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5641 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     5701 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5761 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5821 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5881 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5941 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     6001 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     6061 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     6121 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     6181 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6241 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     6301 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     6361 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     6421 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     6481 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6541 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6601 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     6661 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     6721 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     6781 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     6841 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     6901 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     6961 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     7021 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     7081 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     7141 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     7201 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     7261 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     7321 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     7381 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     7441 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     7501 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     7561 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     7621 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     7681 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     7741 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     7801 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     7861 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     7921 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7981 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     8041 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     8101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     8161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     8221 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     8281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     8341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     8401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     8461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     8521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     8581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     8641 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     8701 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8761 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     8821 tagcccatat atggagttcc g
//
