LOCUS       sequence-36157-e        4866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4866)
  AUTHORS   Weinberg DE, Nakanishi K, Patel DJ, Bartel DP
  TITLE     The inside-out mechanism of dicers from budding yeasts.
  JOURNAL   Cell. 2011 Jul 22;146(2):262-76.
  PUBMED    21784247
REFERENCE   2  (bases 1 to 4866)
  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: "Cell."; 
            date: "2011-07-22"; volume: "146(2)"; pages: "262-76"
COMMENT     Sequence Label: pRSF-KpDcr1deltaC(K217A)
FEATURES             Location/Qualifiers
     source          1..4866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..50
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             60..77
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             90..383
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             505..516
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1485..1503)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1499..1546
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1776..2591)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1776..2591)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2499..2518
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2592..2683)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2699..3448)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2709..2726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2949..2968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3644..4726)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4688..4707
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4727..4804)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctttaataag gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc
       61 tggtgccgcg cggcagccat atggctagca tgtcggactc agaagtcaat caagaagcta
      121 agccagaggt caagccagaa gtcaagcctg agactcacat caatttaaag gtgtccgatg
      181 gatcttcaga gatcttcttc aagatcaaaa agaccactcc tttaagaagg ctgatggaag
      241 cgttcgctaa aagacagggt aaggaaatgg actccttaag attcttgtac gacggtatta
      301 gaatccaagc tgatcagacc cctgaagatt tggacatgga ggataacgat attattgagg
      361 ctcacagaga acagattggt ggatccagca acgagttaaa agttagagaa ttttatagac
      421 tccacaatgc ttgtgtgaaa ttgaaagagt ctattaaatt gatttatgaa aacccattgg
      481 taactgacca aaatgttttg aatttgggaa ctgcagaaaa tactattgat tatacaattt
      541 taaacacacc tacactaaat gttgctaaaa ctttactggg caatagatat tcattagatc
      601 taatagatct ttttcaatct catgatttca aagattcaaa cactgatgtg gatatgttta
      661 ttaaatatcc tgtggtttat gatgaaaatt tggaaaattt agcatttatg cataaaagtt
      721 ttccaaatat gaatgcacat ctaaatgatg ctcaaaagac tcaattatct aatgaaaggt
      781 tagaattttt gggtgatagt tggctaggtg ctttagtttc atatattgta tacaccagat
      841 ttccttctgc aaatgaaggt atgctttctc aaatgaaaga atcaatagta aataataata
      901 atctatttga ttggagtaca aaattaaact tcaccaagag attacaaggt aatatcgcta
      961 cacctactag agtggtcaaa gataaaatgt ctgcacgtta tgctgattgt gttgaagcct
     1021 atattggtgc attggttatt gatagatttg gcactgagtt tttggatatc aaggaatggt
     1081 tagaagaact atcagaaaaa aaattggcca aatcgagcca aatggtgata aaggaaccat
     1141 taaataaaaa tgctaaaaat gaattagccg aattactgca aattaacaaa ttaggacata
     1201 aacttcatta cagaaagctt acagaaatgc ctccattcag agttgaagtt aaaataggtg
     1261 acattctgtt agacgaagct gaaggtaaca gtataaggga agcagaacac agagctgcga
     1321 tgaaggtatt ggaaaatgat gaattattga ggaaatattc ggtttatgat ttagaggatg
     1381 gaaactcgaa gaaatattct attgaagagt aactgctgcg aaatttgaac gccagcacat
     1441 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
     1501 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     1561 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     1621 gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac gaccgggtct
     1681 ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1741 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     1801 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1861 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1921 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1981 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2041 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2101 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcgg
     2161 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2221 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2281 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2341 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2401 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2461 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2521 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     2581 atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2641 atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat gcagcgctct
     2701 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggtgtca
     2761 gctcactcaa aagcggtaat acggttatcc acagaatcag gggataaagc cggaaagaac
     2821 atgtgagcaa aaagcaaagc accggaagaa gccaacgccg caggcgtttt tccataggct
     2881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagc cagaggtggc gaaacccgac
     2941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3061 tcatagctca cgctgttggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccattggta actgatttag
     3241 aggactttgt cttgaagtta tgcacctgtt aaggctaaac tgaaagaaca gattttggtg
     3301 agtgcggtcc tccaacccac ttaccttggt tcaaagagtt ggtagctcag cgaaccttga
     3361 gaaaaccacc gttggtagcg gtggtttttc tttatttatg agatgatgaa tcaatcggtc
     3421 tatcaagtca acgaacagct attccgttac tctagatttc agtgcaattt atctcttcaa
     3481 atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc
     3541 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     3601 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     3661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3721 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     3781 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     3841 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     3901 tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg
     3961 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     4021 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     4081 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     4141 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     4201 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     4261 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     4321 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     4381 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     4441 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     4501 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     4561 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     4621 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     4681 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     4741 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     4801 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaatt aatacgactc
     4861 actata
//
