LOCUS       sequence-36161-e        4305 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 4305)
  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 4305)
  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-KpDcr1deltaNdeltaC
FEATURES             Location/Qualifiers
     source          1..4305
                     /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"
     primer_bind     complement(924..942)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      938..985
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1215..2030)
                     /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(1215..2030)
                     /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     1938..1957
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2031..2122)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2138..2887)
                     /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(2148..2165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2388..2407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3083..4165)
                     /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     4127..4146
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4166..4243)
                     /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 ggatccaaat atcctgtggt ttatgatgaa aatttggaaa
      421 atttagcatt tatgcataaa agttttccaa atatgaatgc acatctaaat gatgctcaaa
      481 agactcaatt atctaatgaa aggttagaat ttttgggtga tagttggcta ggtgctttag
      541 tttcatatat tgtatacacc agatttcctt ctgcaaatga aggtatgctt tctcaaatga
      601 aagaatcaat agtaaataat aataatctat ttgattggag tacaaaatta aacttcacca
      661 agagattaca aggtaatatc gctacaccta ctagagtggt caaagataaa atgtctaaac
      721 gttatgctga ttgtgttgaa gcctatattg gtgcattggt tattgataga tttggcactg
      781 agtttttgga tatcaaggaa tggttagaag aactatcaga aaaaaaattg gccaaatcga
      841 gccaaatgta actgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc
      901 ttaattaacc taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      961 taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac
     1021 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
     1081 accctgccct gaaccgacga caagctgacg accgggtctc cgcaagtggc acttttcggg
     1141 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1201 tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     1261 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     1321 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     1381 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     1441 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     1501 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     1561 ttcattcgtg attgcgcctg agcgagacga aatacgcggt cgctgttaaa aggacaatta
     1621 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     1681 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     1741 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     1801 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     1861 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     1921 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     1981 gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat actcttcctt
     2041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2101 tgtatttaga aaaataaaca aataggcatg cagcgctctt ccgcttcctc gctcactgac
     2161 tcgctacgct cggtcgttcg actgcggcga gcggtgtcag ctcactcaaa agcggtaata
     2221 cggttatcca cagaatcagg ggataaagcc ggaaagaaca tgtgagcaaa aagcaaagca
     2281 ccggaagaag ccaacgccgc aggcgttttt ccataggctc cgcccccctg acgagcatca
     2341 caaaaatcga cgctcaagcc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgttggta
     2521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2641 cttatcgcca ctggcagcag ccattggtaa ctgatttaga ggactttgtc ttgaagttat
     2701 gcacctgtta aggctaaact gaaagaacag attttggtga gtgcggtcct ccaacccact
     2761 taccttggtt caaagagttg gtagctcagc gaaccttgag aaaaccaccg ttggtagcgg
     2821 tggtttttct ttatttatga gatgatgaat caatcggtct atcaagtcaa cgaacagcta
     2881 ttccgttact ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     2941 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     3001 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3061 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3121 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3181 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3241 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3301 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3361 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3421 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     3481 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     3541 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     3601 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     3661 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     3721 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     3781 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     3841 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     3901 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     3961 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4021 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4081 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4141 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4201 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     4261 cccttatgcg actcctgcat taggaaatta atacgactca ctata
//
