LOCUS       sequence-36153-e        5589 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 5589)
  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 5589)
  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
FEATURES             Location/Qualifiers
     source          1..5589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..127
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             140..433
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             597..608
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2126..2170
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2208..2226)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2222..2269
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2499..3314)
                     /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(2499..3314)
                     /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     3222..3241
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3315..3406)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3422..4171)
                     /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(3432..3449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3672..3691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4367..5449)
                     /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     5411..5430
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5450..5527)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5573..5589,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5573..5589,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg
      121 cggcagccat atggctagca tgtcggactc agaagtcaat caagaagcta agccagaggt
      181 caagccagaa gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga
      241 gatcttcttc aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa
      301 aagacagggt aaggaaatgg actccttaag attcttgtac gacggtatta gaatccaagc
      361 tgatcagacc cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga
      421 acagattggt ggatccatgt ctgatcaagt ttatgttccg ccaacatcag caccaatgag
      481 caacgagtta aaagttagag aattttatag actccacaat gcttgtgtga aattgaaaga
      541 gtctattaaa ttgatttatg aaaacccatt ggtaactgac caaaatgttt tgaatttggg
      601 aactgcagaa aatactattg attatacaat tttaaacaca cctacactaa atgttgctaa
      661 aactttactg ggcaatagat attcattaga tctaatagat ctttttcaat ctcatgattt
      721 caaagattca aacactgatg tggatatgtt tattaaatat cctgtggttt atgatgaaaa
      781 tttggaaaat ttagcattta tgcataaaag ttttccaaat atgaatgcac atctaaatga
      841 tgctcaaaag actcaattat ctaatgaaag gttagaattt ttgggtgata gttggctagg
      901 tgctttagtt tcatatattg tatacaccag atttccttct gcaaatgaag gtatgctttc
      961 tcaaatgaaa gaatcaatag taaataataa taatctattt gattggagta caaaattaaa
     1021 cttcaccaag agattacaag gtaatatcgc tacacctact agagtggtca aagataaaat
     1081 gtctaaacgt tatgctgatt gtgttgaagc ctatattggt gcattggtta ttgatagatt
     1141 tggcactgag tttttggata tcaaggaatg gttagaagaa ctatcagaaa aaaaattggc
     1201 caaatcgagc caaatggtga taaaggaacc attaaataaa aatgctaaaa atgaattagc
     1261 cgaattactg caaattaaca aattaggaca taaacttcat tacagaaagc ttacagaaat
     1321 gcctccattc agagttgaag ttaaaatagg tgacattctg ttagacgaag ctgaaggtaa
     1381 cagtataagg gaagcagaac acagagctgc gatgaaggta ttggaaaatg atgaattatt
     1441 gaggaaatat tcggtttatg atttagagga tggaaactcg aagaaatatt ctattgaaga
     1501 gacggatctt aaacatgata ttgaatcctc tggaatatct aatgaaaatt ttttgcaaaa
     1561 tgagggaaat ggaactgaaa atatagaatc aaaatttaaa caattgaata tatcaaatga
     1621 tagcaacaaa gatgatttag ttaataatat tacaaattct gtttttagta aattgcagga
     1681 tgtcgtaagt aaaattgtca ttgacgctat ttccgaatca atcccaaata atccccacaa
     1741 aatagaacaa tctagaaatg gtaaagtagg aacacaaaca aatactagtg ctacaacttc
     1801 agatgtcctt tcagttcaaa cgtctattcc taaaagcgag tcaacttcgg ctgataaagc
     1861 tcctatcaca tgtgatccca atgccaaaca ggcattgtat accctgttat ctagatcaca
     1921 agtgttccca gactatgaaa tggttcaaca ttccatggat aatttcaccg ctaactgtat
     1981 cattaaaggc agcggtatat ttttagcgtc cggaagaggt cgcagtaaga agattgctca
     2041 gcaaaatgca gcatcagcgg cacttcggag cccaatctta aaatctattt tggaagaaca
     2101 aaacaatttt taactcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca
     2161 catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata
     2221 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
     2281 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     2341 atagacataa gcggctattt aacgaccctg ccctgaaccg acgacaagct gacgaccggg
     2401 tctccgcaag tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2461 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     2521 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     2581 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     2641 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     2701 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     2761 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     2821 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     2881 cggtcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     2941 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     3001 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     3061 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     3121 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     3181 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     3241 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     3301 tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3361 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg catgcagcgc
     3421 tcttccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg
     3481 tcagctcact caaaagcggt aatacggtta tccacagaat caggggataa agccggaaag
     3541 aacatgtgag caaaaagcaa agcaccggaa gaagccaacg ccgcaggcgt ttttccatag
     3601 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc
     3661 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3721 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3781 ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3841 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3901 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccattg gtaactgatt
     3961 tagaggactt tgtcttgaag ttatgcacct gttaaggcta aactgaaaga acagattttg
     4021 gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga gttggtagct cagcgaacct
     4081 tgagaaaacc accgttggta gcggtggttt ttctttattt atgagatgat gaatcaatcg
     4141 gtctatcaag tcaacgaaca gctattccgt tactctagat ttcagtgcaa tttatctctt
     4201 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat
     4261 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     4321 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     4381 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4441 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     4501 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     4561 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     4621 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
     4681 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     4741 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     4801 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     4861 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     4921 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     4981 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     5041 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     5101 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     5161 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     5221 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     5281 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     5341 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     5401 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     5461 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     5521 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga
     5581 ctcactata
//
