LOCUS       sequence-36162-e        4995 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 4995)
  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 4995)
  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-KpDcr1(1-384)
FEATURES             Location/Qualifiers
     source          1..4995
                     /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             547..558
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1614..1632)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1628..1675
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1905..2720)
                     /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(1905..2720)
                     /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     2628..2647
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2721..2812)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2828..3577)
                     /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(2838..2855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3078..3097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3773..4855)
                     /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     4817..4836
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4856..4933)
                     /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 ggatccatgt ctgatcaagt ttatgttccg ccaacatcag
      421 caccaatgag caacgagtta aaagttagag aattttatag actccacaat gcttgtgtga
      481 aattgaaaga gtctattaaa ttgatttatg aaaacccatt ggtaactgac caaaatgttt
      541 tgaatttggg aactgcagaa aatactattg attatacaat tttaaacaca cctacactaa
      601 atgttgctaa aactttactg ggcaatagat attcattaga tctaatagat ctttttcaat
      661 ctcatgattt caaagattca aacactgatg tggatatgtt tattaaatat cctgtggttt
      721 atgatgaaaa tttggaaaat ttagcattta tgcataaaag ttttccaaat atgaatgcac
      781 atctaaatga tgctcaaaag actcaattat ctaatgaaag gttagaattt ttgggtgata
      841 gttggctagg tgctttagtt tcatatattg tatacaccag atttccttct gcaaatgaag
      901 gtatgctttc tcaaatgaaa gaatcaatag taaataataa taatctattt gattggagta
      961 caaaattaaa cttcaccaag agattacaag gtaatatcgc tacacctact agagtggtca
     1021 aagataaaat gtctaaacgt tatgctgatt gtgttgaagc ctatattggt gcattggtta
     1081 ttgatagatt tggcactgag tttttggata tcaaggaatg gttagaagaa ctatcagaaa
     1141 aaaaattggc caaatcgagc caaatggtga taaaggaacc attaaataaa aatgctaaaa
     1201 atgaattagc cgaattactg caaattaaca aattaggaca taaacttcat tacagaaagc
     1261 ttacagaaat gcctccattc agagttgaag ttaaaatagg tgacattctg ttagacgaag
     1321 ctgaaggtaa cagtataagg gaagcagaac acagagctgc gatgaaggta ttggaaaatg
     1381 atgaattatt gaggaaatat tcggtttatg atttagagga tggaaactcg aagaaatatt
     1441 ctattgaaga gacggatctt aaacatgata ttgaatcctc tggaatatct aatgaaaatt
     1501 ttttgcaaaa tgagggaaat ggaactgaaa atatagaata actgctgcga aatttgaacg
     1561 ccagcacatg gactcgtcta ctagcgcagc ttaattaacc taggctgctg ccaccgctga
     1621 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1681 acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     1741 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga caagctgacg
     1801 accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1861 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
     1921 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     1981 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     2041 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     2101 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     2161 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     2221 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     2281 aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     2341 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     2401 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     2461 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     2521 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     2581 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     2641 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     2701 tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2761 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggcatg
     2821 cagcgctctt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     2881 gcggtgtcag ctcactcaaa agcggtaata cggttatcca cagaatcagg ggataaagcc
     2941 ggaaagaaca tgtgagcaaa aagcaaagca ccggaagaag ccaacgccgc aggcgttttt
     3001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagcc agaggtggcg
     3061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3181 ggcgctttct catagctcac gctgttggta tctcagttcg gtgtaggtcg ttcgctccaa
     3241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccattggtaa
     3361 ctgatttaga ggactttgtc ttgaagttat gcacctgtta aggctaaact gaaagaacag
     3421 attttggtga gtgcggtcct ccaacccact taccttggtt caaagagttg gtagctcagc
     3481 gaaccttgag aaaaccaccg ttggtagcgg tggtttttct ttatttatga gatgatgaat
     3541 caatcggtct atcaagtcaa cgaacagcta ttccgttact ctagatttca gtgcaattta
     3601 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
     3661 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     3721 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3841 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     3901 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     3961 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     4021 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     4081 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     4141 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     4201 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     4261 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     4321 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     4381 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     4441 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     4501 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     4561 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     4621 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     4681 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     4741 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     4801 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     4861 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4921 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaatta
     4981 atacgactca ctata
//
