LOCUS       sequence-193803-       10532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of helicase mutant of human Dicer in Sf9 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10532)
  AUTHORS   MacRae IJ, Ma E, Zhou M, Robinson CV, Doudna JA
  TITLE     In vitro reconstitution of the human RISC-loading complex.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Jan 15;105(2):512-7. doi: 
            10.1073/pnas.0710869105. Epub 2008 Jan 4.
  PUBMED    18178619
REFERENCE   2  (bases 1 to 10532)
  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: 
            "10.1073/pnas.0710869105"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2008-01-15- 15"; volume: "105"; issue: "2"; pages: "512-7"
COMMENT     Sequence Label: pFastBac-hDcr-K70A.
FEATURES             Location/Qualifiers
     source          1..10532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        179..270
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     195..212
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             337..354
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             376..396
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    6316..6450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6353..6372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6407..6426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6479..6644
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6828..7282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6915..6934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7124..7145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7308..7412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7413..7481,7482..8273)
                     /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     complement(7631..7650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8444..9032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8933..8952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9186..9203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  9337..9561
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(9628..10161)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(10331..10336,10337..10353,10354..10359))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg ccggtggtgc
       61 tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg
      121 cccaggactc tagctatagt tctagtggtt ggctacgtat actccggaat attaatagat
      181 catggagata attaaaatga taaccatctc gcaaataaat aagtatttta ctgttttcgt
      241 aacagttttg taataaaaaa acctataaat attccggatt attcataccg tcccaccatc
      301 gggcgcggat ctcggtccga aaccatgtcg tactaccatc accatcacca tcacgattac
      361 gatatcccaa cgaccgaaaa cctgtatttt cagggcgcca tgaaaagccc tgctttgcaa
      421 cccctcagca tggcaggcct gcagctcatg acccctgctt cctcaccaat gggtcctttc
      481 tttggactgc catggcaaca agaagcaatt catgataaca tttatacgcc aagaaaatat
      541 caggttgaac tgcttgaagc agctctggat cataatacca tcgtctgttt aaacactggc
      601 tcaggggcga catttattgc agtactactc actaaagagc tgtcctatca gatcagggga
      661 gacttcagca gaaatggaaa aaggacggtg ttcttggtca actctgcaaa ccaggttgct
      721 caacaagtgt cagctgtcag aactcattca gatctcaagg ttggggaata ctcaaaccta
      781 gaagtaaatg catcttggac aaaagagaga tggaaccaag agtttactaa gcaccaggtt
      841 ctcattatga cttgctatgt cgccttgaat gttttgaaaa atggttactt atcactgtca
      901 gacattaacc ttttggtgtt tgatgagtgt catcttgcaa tcctagacca cccctatcga
      961 gaaattatga agctctgtga aaattgtcca tcatgtcctc gcattttggg actaactgct
     1021 tccattttaa atgggaaatg tgatccagag gaattggaag aaaagattca gaaactagag
     1081 aaaattctta agagtaatgc tgaaactgca actgacctgg tggtcttaga caggtatact
     1141 tctcagccat gtgagattgt ggtggattgt ggaccattta ctgacagaag tgggctttat
     1201 gaaagactgc tgatggaatt agaagaagca cttaatttta tcaatgattg taatatatct
     1261 gtacattcaa aagaaagaga ttctacttta atttcgaaac agatactatc agactgtcgt
     1321 gccgtattgg tagttctggg accctggtgt gcagataaag tagctggaat gatggtaaga
     1381 gaactacaga aatacatcaa acatgagcaa gaggagctgc acaggaaatt tttattgttt
     1441 acagacactt tcctaaggaa aatacatgca ctatgtgaag agcacttctc acctgcctca
     1501 cttgacctga aatttgtaac tcctaaagta atcaaactgc tcgaaatctt acgcaaatat
     1561 aaaccatatg agcgacagca gtttgaaagc gttgagtggt ataataatag aaatcaggat
     1621 aattatgtgt catggagtga ttctgaggat gatgatgagg atgaagaaat tgaagaaaaa
     1681 gagaagccag agacaaattt tccttctcct tttaccaaca ttttgtgcgg aattattttt
     1741 gtggaaagaa gatacacagc agttgtctta aacagattga taaaggaagc tggcaaacaa
     1801 gatccagagc tggcttatat cagtagcaat ttcataactg gacatggcat tgggaagaat
     1861 cagcctcgca acaaacagat ggaagcagaa ttcagaaaac aggaagaggt acttaggaaa
     1921 tttcgagcac atgagaccaa cctgcttatt gcaacaagta ttgtagaaga gggtgttgat
     1981 ataccaaaat gcaacttggt ggttcgtttt gatttgccca cagaatatcg atcctatgtt
     2041 caatctaaag gaagagcaag ggcacccatc tctaattata taatgttagc ggatacagac
     2101 aaaataaaaa gttttgaaga agaccttaaa acctacaaag ctattgaaaa gatcttgaga
     2161 aacaagtgtt ccaagtcggt tgatactggt gagactgaca ttgatcctgt catggatgat
     2221 gatgacgttt tcccaccata tgtgttgagg cctgacgatg gtggtccacg agtcacaatc
     2281 aacacggcca ttggacacat caatagatac tgtgctagat taccaagtga tccgtttact
     2341 catctagctc ctaaatgcag aacccgagag ttgcctgatg gtacatttta ttcaactctt
     2401 tatctgccaa ttaactcacc tcttcgagcc tccattgttg gtccaccaat gagctgtgta
     2461 cgattggctg aaagagttgt agctctcatt tgctgtgaga aactgcacaa aattggcgaa
     2521 ctggatgacc atttgatgcc agttgggaaa gagactgtta aatatgaaga ggagcttgat
     2581 ttgcatgatg aagaagagac cagtgttcca ggaagaccag gttccacgaa acgaaggcag
     2641 tgctacccaa aagcaattcc agagtgtttg agggatagtt atcccagacc tgatcagccc
     2701 tgttacctgt atgtgatagg aatggtttta actacacctt tacctgatga actcaacttt
     2761 agaaggcgga agctctatcc tcctgaagat accacaagat gctttggaat actgacggcc
     2821 aaacccatac ctcagattcc acactttcct gtgtacacac gctctggaga ggttaccata
     2881 tccattgagt tgaagaagtc tggtttcatg ttgtctctac aaatgcttga gttgattaca
     2941 agacttcacc agtatatatt ctcacatatt cttcggcttg aaaaacctgc actagaattt
     3001 aaacctacag acgctgattc agcatactgt gttctacctc ttaatgttgt taatgactcc
     3061 agcactttgg atattgactt taaattcatg gaagatattg agaagtctga agctcgcata
     3121 ggcattccca gtacaaagta tacaaaagaa acaccctttg tttttaaatt agaagattac
     3181 caagatgccg ttatcattcc aagatatcgc aattttgatc agcctcatcg attttatgta
     3241 gctgatgtgt acactgatct taccccactc agtaaatttc cttcccctga gtatgaaact
     3301 tttgcagaat attataaaac aaagtacaac cttgacctaa ccaatctcaa ccagccactg
     3361 ctggatgtgg accacacatc ttcaagactt aatcttttga cacctcgaca tttgaatcag
     3421 aaggggaaag cgcttccttt aagcagtgct gagaagagga aagccaaatg ggaaagtctg
     3481 cagaataaac agatactggt tccagaactc tgtgctatac atccaattcc agcatcactg
     3541 tggagaaaag ctgtttgtct ccccagcata ctttatcgcc ttcactgcct tttgactgca
     3601 gaggagctaa gagcccagac tgccagcgat gctggcgtgg gagtcagatc acttcctgcg
     3661 gattttagat accctaactt agacttcggg tggaaaaaat ctattgacag caaatctttc
     3721 atctcaattt ctaactcctc ttcagctgaa aatgataatt actgtaagca cagcacaatt
     3781 gtccctgaaa atgctgcaca tcaaggtgct aatagaacct cctctctaga aaatcatgac
     3841 caaatgtctg tgaactgcag aacgttgctc agcgagtccc ctggtaagct ccacgttgaa
     3901 gtttcagcag atcttacagc aattaatggt ctttcttaca atcaaaatct cgccaatggc
     3961 agctatgatt tagctaacag agacttttgc caaggaaatc agctaaatta ctacaagcag
     4021 gaaatacccg tgcaaccaac tacctcatat tccattcaga atttatacag ttacgagaac
     4081 cagccccagc ccagcgatga atgtactctc ctgagtaata aataccttga tggaaatgct
     4141 aacaaatcta cctcagatgg aagtcctgtg atggccgtaa tgcctggtac gacagacact
     4201 attcaagtgc tcaagggcag gatggattct gagcagagcc cttctattgg gtactcctca
     4261 aggactcttg gccccaatcc tggacttatt cttcaggctt tgactctgtc aaacgctagt
     4321 gatggattta acctggagcg gcttgaaatg cttggcgact cctttttaaa gcatgccatc
     4381 accacatatc tattttgcac ttaccctgat gcgcatgagg gccgcctttc atatatgaga
     4441 agcaaaaagg tcagcaactg taatctgtat cgccttggaa aaaagaaggg actacccagc
     4501 cgcatggtgg tgtcaatatt tgatccccct gtgaattggc ttcctcctgg ttatgtagta
     4561 aatcaagaca aaagcaacac agataaatgg gaaaaagatg aaatgacaaa agactgcatg
     4621 ctggcgaatg gcaaactgga tgaggattac gaggaggagg atgaggagga ggagagcctg
     4681 atgtggaggg ctccgaagga agaggctgac tatgaagatg atttcctgga gtatgatcag
     4741 gaacatatca gatttataga taatatgtta atggggtcag gagcttttgc aaagaaaatc
     4801 tctctttctc ctttttcaac cactgattct gcatatgaat ggaaaatgcc caaaaaatcc
     4861 tccttaggta gtatgccatt ttcatcagat tttgaggatt ttgactacag ctcttgggat
     4921 gcaatgtgct atctggatcc tagcaaagct gttgaagaag atgactttgt ggtggggttc
     4981 tggaatccat cagaagaaaa ctgtggtgtt gacacgggaa agcagtccat ttcttacgac
     5041 ttgcacactg agcagtgtat tgctgacaaa agcatagcgg actgtgtgga agccctgctg
     5101 ggctgctatt taaccagctg tggggagagg gctgctcagc ttttcctctg ttcactgggg
     5161 ctgaaggtgc tcccggtaat taaaaggact gatcgggaaa aggccctgtg ccctactcgg
     5221 gagaatttca acagccaaca aaagaacctt tcagtgagct gtgctgctgc ttctgtggcc
     5281 agttcacgct cttctgtatt gaaagactcg gaatatggtt gtttgaagat tccaccaaga
     5341 tgtatgtttg atcatccaga tgcagataaa acactgaatc accttatatc ggggtttgaa
     5401 aattttgaaa agaaaatcaa ctacagattc aagaataagg cttaccttct ccaggctttt
     5461 acacatgcct cctaccacta caatactatc actgattgtt accagcgctt agaattcctg
     5521 ggagatgcga ttttggacta cctcataacc aagcaccttt atgaagaccc gcggcagcac
     5581 tccccggggg tcctgacaga cctgcggtct gccctggtca acaacaccat ctttgcatcg
     5641 ctggctgtaa agtacgacta ccacaagtac ttcaaagctg tctctcctga gctcttccat
     5701 gtcattgatg actttgtgca gtttcagctt gagaagaatg aaatgcaagg aatggattct
     5761 gagcttagga gatctgagga ggatgaagag aaagaagagg atattgaagt tccaaaggcc
     5821 atgggggata tttttgagtc gcttgctggt gccatttaca tggatagtgg gatgtcactg
     5881 gagacagtct ggcaggtgta ctatcccatg atgcggccac taatagaaaa gttttctgca
     5941 aatgtacccc gttcccctgt gcgagaattg cttgaaatgg aaccagaaac tgccaaattt
     6001 agcccggctg agagaactta cgacgggaag gtcagagtca ctgtggaagt agtaggaaag
     6061 gggaaattta aaggtgttgg tcgaagttac aggattgcca aatctgcagc agcaagaaga
     6121 gccctccgaa gcctcaaagc taatcaacct caggttccca atagctgact cgaggcatgc
     6181 ggtaccaagc ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag
     6241 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     6301 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6361 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6421 ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa
     6481 ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta
     6541 cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact
     6601 ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt
     6661 cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc
     6721 tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt
     6781 aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag
     6841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6901 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6961 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7021 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7081 gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa
     7141 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7201 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7261 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     7381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     7441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7621 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     7681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7921 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     8041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     8101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     8161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     8221 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     8281 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     8341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     8401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8581 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     9001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     9061 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     9121 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     9181 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     9241 cgcatctgtg cggtatttca caccgcatag accagccgcg taacctggca aaatcggtta
     9301 cggttgagta ataaatggat gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa
     9361 actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag aatagttgta
     9421 aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag
     9481 caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggccaag
     9541 ggcatggtaa agactatatt cgcggcgttg tgacaattta ccgaacaact ccgcggccgg
     9601 gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg
     9661 catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta
     9721 atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt
     9781 gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat
     9841 atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac
     9901 aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc
     9961 gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc
    10021 gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg
    10081 aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac
    10141 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
    10201 tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
    10261 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
    10321 cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg
    10381 tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc
    10441 ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt
    10501 gctgttcttc tacggcaagg tgctgtgcac gg
//
