LOCUS       sequence-200866-        9655 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9655)
  AUTHORS   Gurtan AM, Lu V, Bhutkar A, Sharp PA
  TITLE     In vivo structure-function analysis of human Dicer reveals 
            directional processing of precursor miRNAs.
  JOURNAL   RNA. 2012 Jun;18(6):1116-22. Epub 2012 Apr 30.
  PUBMED    22546613
REFERENCE   2  (bases 1 to 9655)
  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"; journalName: "RNA."; 
            date: "2012-06"; volume: "18(6)"; pages: "1116-22. Epub 2012 Apr 30"
COMMENT     Sequence Label: pCAGGS-Flag-hsDicer
FEATURES             Location/Qualifiers
     source          1..9655
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     340..362
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     424..443
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             490..513
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6292..6311)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6357..6412
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6411..6430)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6773..6789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6773..6789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6786..6808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6797..6813
                     /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(6821..6827,6828..6845,6846..6851))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6866..6887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6946..7141
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6992..7127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7147..7281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7184..7203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7238..7257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7349..7366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7520..8108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7600..7619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8279..9070,9071..9139))
                     /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     8902..8921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9140..9244)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        9275..9654
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
       61 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
      121 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
      181 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
      241 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
      301 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
      361 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
      421 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc tcgagattga
      481 tctaccatgg actacaaaga cgatgacgac aagcttaaaa gccctgcttt gcaacccctc
      541 agcatggcag gcctgcagct catgacccct gcttcctcac caatgggtcc tttctttgga
      601 ctgccatggc aacaagaagc aattcatgat aacatttata cgccaagaaa atatcaggtt
      661 gaactgcttg aagcagctct ggatcataat accatcgtct gtttaaacac tggctcaggg
      721 aagacattta ttgcagtact actcactaaa gagctgtcct atcagatcag gggagacttc
      781 agcagaaatg gaaaaaggac ggtgttcttg gtcaactctg caaaccaggt tgctcaacaa
      841 gtgtcagctg tcagaactca ttcagatctc aaggttgggg aatactcaaa cctagaagta
      901 aatgcatctt ggacaaaaga gagatggaac caagagttta ctaagcacca ggttctcatt
      961 atgacttgct atgtcgcctt gaatgttttg aaaaatggtt acttatcact gtcagacatt
     1021 aaccttttgg tgtttgatga gtgtcatctt gcaatcctag accaccccta tcgagaaatt
     1081 atgaagctct gtgaaaattg tccatcatgt cctcgcattt tgggactaac tgcttccatt
     1141 ttaaatggga aatgtgatcc agaggaattg gaagaaaaga ttcagaaact agagaaaatt
     1201 cttaagagta atgctgaaac tgcaactgac ctggtggtct tagacaggta tacttctcag
     1261 ccatgtgaga ttgtggtgga ttgtggacca tttactgaca gaagtgggct ttatgaaaga
     1321 ctgctgatgg aattagaaga agcacttaat tttatcaatg attgtaatat atctgtacat
     1381 tcaaaagaaa gagattctac tttaatttcg aaacagatac tatcagactg tcgtgccgta
     1441 ttggtagttc tgggaccctg gtgtgcagat aaagtagctg gaatgatggt aagagaacta
     1501 cagaaataca tcaaacatga gcaagaggag ctgcacagga aatttttatt gtttacagac
     1561 actttcctaa ggaaaataca tgcactatgt gaagagcact tctcacctgc ctcacttgac
     1621 ctgaaatttg taactcctaa agtaatcaaa ctgctcgaaa tcttacgcaa atataaacca
     1681 tatgagcgac agcagtttga aagcgttgag tggtataata atagaaatca ggataattat
     1741 gtgtcatgga gtgattctga ggatgatgat gaggatgaag aaattgaaga aaaagagaag
     1801 ccagagacaa attttccttc tccttttacc aacattttgt gcggaattat ttttgtggaa
     1861 agaagataca cagcagttgt cttaaacaga ttgataaagg aagctggcaa acaagatcca
     1921 gagctggctt atatcagtag caatttcata actggacatg gcattgggaa gaatcagcct
     1981 cgcaacaaac agatggaagc agaattcaga aaacaggaag aggtacttag gaaatttcga
     2041 gcacatgaga ccaacctgct tattgcaaca agtattgtag aagagggtgt tgatatacca
     2101 aaatgcaact tggtggttcg ttttgatttg cccacagaat atcgatccta tgttcaatct
     2161 aaaggaagag caagggcacc catctctaat tatataatgt tagcggatac agacaaaata
     2221 aaaagttttg aagaagacct taaaacctac aaagctattg aaaagatctt gagaaacaag
     2281 tgttccaagt cggttgatac tggtgagact gacattgatc ctgtcatgga tgatgatgac
     2341 gttttcccac catatgtgtt gaggcctgac gatggtggtc cacgagtcac aatcaacacg
     2401 gccattggac acatcaatag atactgtgct agattaccaa gtgatccgtt tactcatcta
     2461 gctcctaaat gcagaacccg agagttgcct gatggtacat tttattcaac tctttatctg
     2521 ccaattaact cacctcttcg agcctccatt gttggtccac caatgagctg tgtacgattg
     2581 gctgaaagag ttgtagctct catttgctgt gagaaactgc acaaaattgg cgaactggat
     2641 gaccatttga tgccagttgg gaaagagact gttaaatatg aagaggagct tgatttgcat
     2701 gatgaagaag agaccagtgt tccaggaaga ccaggttcca cgaaacgaag gcagtgctac
     2761 ccaaaagcaa ttccagagtg tttgagggat agttatccca gacctgatca gccctgttac
     2821 ctgtatgtga taggaatggt tttaactaca cctttacctg atgaactcaa ctttagaagg
     2881 cggaagctct atcctcctga agataccaca agatgctttg gaatactgac ggccaaaccc
     2941 atacctcaga ttccacactt tcctgtgtac acacgctctg gagaggttac catatccatt
     3001 gagttgaaga agtctggttt catgttgtct ctacaaatgc ttgagttgat tacaagactt
     3061 caccagtata tattctcaca tattcttcgg cttgaaaaac ctgcactaga atttaaacct
     3121 acagacgctg attcagcata ctgtgttcta cctcttaatg ttgttaatga ctccagcact
     3181 ttggatattg actttaaatt catggaagat attgagaagt ctgaagctcg cataggcatt
     3241 cccagtacaa agtatacaaa agaaacaccc tttgttttta aattagaaga ttaccaagat
     3301 gccgttatca ttccaagata tcgcaatttt gatcagcctc atcgatttta tgtagctgat
     3361 gtgtacactg atcttacccc actcagtaaa tttccttccc ctgagtatga aacttttgca
     3421 gaatattata aaacaaagta caaccttgac ctaaccaatc tcaaccagcc actgctggat
     3481 gtggaccaca catcttcaag acttaatctt ttgacacctc gacatttgaa tcagaagggg
     3541 aaagcgcttc ctttaagcag tgctgagaag aggaaagcca aatgggaaag tctgcagaat
     3601 aaacagatac tggttccaga actctgtgct atacatccaa ttccagcatc actgtggaga
     3661 aaagctgttt gtctccccag catactttat cgccttcact gccttttgac tgcagaggag
     3721 ctaagagccc agactgccag cgatgctggc gtgggagtca gatcacttcc tgcggatttt
     3781 agatacccta acttagactt cgggtggaaa aaatctattg acagcaaatc tttcatctca
     3841 atttctaact cctcttcagc tgaaaatgat aattactgta agcacagcac aattgtccct
     3901 gaaaatgctg cacatcaagg tgctaataga acctcctctc tagaaaatca tgaccaaatg
     3961 tctgtgaact gcagaacgtt gctcagcgag tcccctggta agctccacgt tgaagtttca
     4021 gcagatctta cagcaattaa tggtctttct tacaatcaaa atctcgccaa tggcagttat
     4081 gatttagcta acagagactt ttgccaagga aatcagctaa attactacaa gcaggaaata
     4141 cccgtgcaac caactacctc atattccatt cagaatttat acagttacga gaaccagccc
     4201 cagcccagcg atgaatgtac tctcctgagt aataaatacc ttgatggaaa tgctaacaaa
     4261 tctacctcag atggaagtcc tgtgatggcc gtaatgcctg gtacgacaga cactattcaa
     4321 gtgctcaagg gcaggatgga ttctgagcag agcccttcta ttgggtactc ctcaaggact
     4381 cttggcccca atcctggact tattcttcag gctttgactc tgtcaaacgc tagtgatgga
     4441 tttaacctgg agcggcttga aatgcttggc gactcctttt taaagcatgc catcaccaca
     4501 tatctatttt gcacttaccc tgatgcgcat gagggccgcc tttcatatat gagaagcaaa
     4561 aaggtcagca actgtaatct gtatcgcctt ggaaaaaaga agggactacc cagccgcatg
     4621 gtggtgtcaa tatttgatcc ccctgtgaat tggcttcctc ctggttatgt agtaaatcaa
     4681 gacaaaagca acacagataa atgggaaaaa gatgaaatga caaaagactg catgctggcg
     4741 aatggcaaac tggatgagga ttacgaggag gaggatgagg aggaggagag cctgatgtgg
     4801 agggctccga aggaagaggc tgactatgaa gatgatttcc tggagtatga tcaggaacat
     4861 atcagattta tagataatat gttaatgggg tcaggagctt ttgtaaagaa aatctctctt
     4921 tctccttttt caaccactga ttctgcatat gaatggaaaa tgcccaaaaa atcctcctta
     4981 ggtagtatgc cattttcatc agattttgag gattttgact acagctcttg ggatgcaatg
     5041 tgctatctgg atcctagcaa agctgttgaa gaagatgact ttgtggtggg gttctggaat
     5101 ccatcagaag aaaactgtgg tgttgacacg ggaaagcagt ccatttctta cgacttgcac
     5161 actgagcagt gtattgctga caaaagcata gcggactgtg tggaagccct gctgggctgc
     5221 tatttaacca gctgtgggga gagggctgct cagcttttcc tctgttcact ggggctgaag
     5281 gtgctcccgg taattaaaag gactgatcgg gaaaaggccc tgtgccctac tcgggagaat
     5341 ttcaacagcc aacaaaagaa cctttcagtg agctgtgctg ctgcttctgt ggccagttca
     5401 cgctcttctg tattgaaaga ctcggaatat ggttgtttga agattccacc aagatgtatg
     5461 tttgatcatc cagatgcaga taaaacactg aatcacctta tatcggggtt tgaaaatttt
     5521 gaaaagaaaa tcaactacag attcaagaat aaggcttacc ttctccaggc ttttacacat
     5581 gcctcctacc actacaatac tatcactgat tgttaccagc gcttagaatt cctgggagat
     5641 gcgattttgg actacctcat aaccaagcac ctttatgaag acccgcggca gcactccccg
     5701 ggggtcctga cagacctgcg gtctgccctg gtcaacaaca ccatctttgc atcgctggct
     5761 gtaaagtacg actaccacaa gtacttcaaa gctgtctctc ctgagctctt ccatgtcatt
     5821 gatgactttg tgcagtttca gcttgagaag aatgaaatgc aaggaatgga ttctgagctt
     5881 aggagatctg aggaggatga agagaaagaa gaggatattg aagttccaaa ggccatgggg
     5941 gatatttttg agtcgcttgc tggtgccatt tacatggata gtgggatgtc actggagaca
     6001 gtctggcagg tgtactatcc catgatgcgg ccactaatag aaaagttttc tgcaaatgta
     6061 ccccgttccc ctgtgcgaga attgcttgaa atggaaccag aaactgccaa atttagcccg
     6121 gctgagagaa cttacgacgg gaaggtcaga gtcactgtgg aagtagtagg aaaggggaaa
     6181 tttaaaggtg ttggtcgaag ttacaggatt gccaaatctg cagcagcaag aagagccctc
     6241 cgaagcctca aagctaatca acctcaggtt cccaatagct gaggtaccag atctttttcc
     6301 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     6361 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     6421 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     6481 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     6541 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     6601 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     6661 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     6721 tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     6781 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6841 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6901 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     6961 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     7021 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     7081 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     7141 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7261 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7321 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7381 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7441 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7501 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7561 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7621 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7681 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7741 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7801 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7861 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7921 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7981 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8041 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8101 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8161 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8221 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8281 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8341 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8401 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8461 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8521 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8581 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8641 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8701 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8761 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8821 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8881 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8941 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9001 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9061 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9121 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9181 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9241 cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagttattaa
     9301 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     9361 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     9421 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     9481 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     9541 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     9601 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgg
//
