LOCUS       sequence-164287-        7761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid, K. polysporus Dcr1 under TEF promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7761)
  AUTHORS   Nakanishi K, Weinberg DE, Bartel DP, Patel DJ
  TITLE     Structure of yeast Argonaute with guide RNA.
  JOURNAL   Nature. 2012 Jun 20;486(7403):368-74. doi: 10.1038/nature11211.
  PUBMED    22722195
REFERENCE   2  (bases 1 to 7761)
  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"; journalName: 
            "Nature"; date: "2012-06-20- 20"; volume: "486"; issue: "7403"; 
            pages: "368-74"
COMMENT     Sequence Label: pRS405-LEU2-TEF-KpolDcr1v2
FEATURES             Location/Qualifiers
     source          1..7761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2241..2261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2279..2677
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2681..2697
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2681..2697
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2859..2870
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      4382..4629
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4398..4416)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4647..4666)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4648..4666)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4676..4693)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4676..4692)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4685..4707)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4833..5288
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4920..4939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5130..5151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5588..5992
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6005..7099
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(7592..7611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7711..7733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctca
     2281 tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat
     2341 cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct
     2401 agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt
     2461 ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt
     2521 ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt
     2581 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
     2641 tgctcattag aaagaaagca tagcaatcta atctaagttt tctagaacta gtggatccat
     2701 gtctgatcaa gtttatgttc cgccaacatc agcaccaatg agcaacgagt taaaagttag
     2761 agaattttat agactccaca atgcttgtgt gaaattgaaa gagtctatta aattgattta
     2821 tgaaaaccca ttggtaactg accaaaatgt tttgaatttg ggaactgcag aaaatactat
     2881 tgattataca attttaaaca cacctacact aaatgttgct aaaactttac tgggcaatag
     2941 atattcatta gatctaatag atctttttca atctcatgat ttcaaagatt caaacactga
     3001 tgtggatatg tttattaaat atcctgtggt ttatgatgaa aatttggaaa atttagcatt
     3061 tatgcataaa agttttccaa atatgaatgc acatctaaat gatgctcaaa agactcaatt
     3121 atctaatgaa aggttagaat ttttgggtga tagttggcta ggtgctttag tttcatatat
     3181 tgtatacacc agatttcctt ctgcaaatga aggtatgctt tctcaaatga aagaatcaat
     3241 agtaaataat aataatctat ttgattggag tacaaaatta aacttcacca agagattaca
     3301 aggtaatatc gctacaccta ctagagtggt caaagataaa atgtctaaac gttatgctga
     3361 ttgtgttgaa gcctatattg gtgcattggt tattgataga tttggcactg agtttttgga
     3421 tatcaaggaa tggttagaag aactatcaga aaaaaaattg gccaaatcga gccaaatggt
     3481 gataaaggaa ccattaaata aaaatgctaa aaatgaatta gccgaattac tgcaaattaa
     3541 caaattagga cataaacttc attacagaaa gcttacagaa atgcctccat tcagagttga
     3601 agttaaaata ggtgacattc tgttagacga agctgaaggt aacagtataa gggaagcaga
     3661 acacagagct gcgatgaagg tattggaaaa tgatgaatta ttgaggaaat attcggttta
     3721 tgatttagag gatggaaact cgaagaaata ttctattgaa gagacggatc ttaaacatga
     3781 tattgaatcc tctggaatat ctaatgaaaa ttttttgcaa aatgagggaa atggaactga
     3841 aaatatagaa tcaaaattta aacaattgaa tatatcaaat gatagcaaca aagatgattt
     3901 agttaataat attacaaatt ctgtttttag taaattgcag gatgtcgtaa gtaaaattgt
     3961 cattgacgct atttccgaat caatcccaaa taatccccac aaaatagaac aatctagaaa
     4021 tggtaaagta ggaacacaaa caaatactag tgctacaact tcagatgtcc tttcagttca
     4081 aacgtctatt cctaaaagcg agtcaacttc ggctgataaa gctcctatca catgtgatcc
     4141 caatgccaaa caggcattgt ataccctgtt atctagatca caagtgttcc cagactatga
     4201 aatggttcaa cattccatgg ataatttcac cgctaactgt atcattaaag gcagcggtat
     4261 atttttagcg tccggaagag gtcgcagtaa gaagattgct cagcaaaatg cagcatcagc
     4321 ggcacttcgg agcccaatct taaaatctat tttggaagaa caaaacaatt tttaactcga
     4381 gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
     4441 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
     4501 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
     4561 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
     4621 ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     4681 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     4741 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     4801 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     4861 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4921 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4981 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5041 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5101 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5161 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5221 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     5281 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5341 tcgacggtcg aggagaactt ctagtatatc cacataccta atattattgc cttattaaaa
     5401 atggaatccc aacaattaca tcaaaatcca cattctcttc aaaatcaatt gtcctgtact
     5461 tccttgttca tgtgtgttca aaaacgttat atttatagga taattatact ctatttctca
     5521 acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct gattcaagaa atatcttgac
     5581 cgcagttaac tgtgggaata ctcaggtatc gtaagatgca agagttcgaa tctcttagca
     5641 accattattt ttttcctcaa cataacgaga acacacaggg gcgctatcgc acagaatcaa
     5701 attcgatgac tggaaatttt ttgttaattt cagaggtcgc ctgacgcata tacctttttc
     5761 aactgaaaaa ttgggagaaa aaggaaaggt gagaggccgg aaccggcttt tcatatagaa
     5821 tagagaagcg ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt
     5881 aaggacctat tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt
     5941 accttttaca tttcagcaat atatatatat atttcaagga tataccattc taatgtctgc
     6001 ccctatgtct gcccctaaga agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat
     6061 cacagccgaa gccattaagg ttcttaaagc tatttctgat gttcgttcca atgtcaagtt
     6121 cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat gctacaggtg tcccacttcc
     6181 agatgaggcg ctggaagcct ccaagaaggt tgatgccgtt ttgttaggtg ctgtggctgg
     6241 tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga
     6301 acttcaattg tacgccaact taagaccatg taactttgca tccgactctc ttttagactt
     6361 atctccaatc aagccacaat ttgctaaagg tactgacttc gttgttgtca gagaattagt
     6421 gggaggtatt tactttggta agagaaagga agacgatggt gatggtgtcg cttgggatag
     6481 tgaacaatac accgttccag aagtgcaaag aatcacaaga atggccgctt tcatggccct
     6541 acaacatgag ccaccattgc ctatttggtc cttggataaa gctaatcttt tggcctcttc
     6601 aagattatgg agaaaaactg tggaggaaac catcaagaac gaattcccta cattgaaggt
     6661 tcaacatcaa ttgattgatt ctgccgccat gatcctagtt aagaacccaa cccacctaaa
     6721 tggtattata atcaccagca acatgtttgg tgatatcatc tccgatgaag cctccgttat
     6781 cccaggttcc ttgggtttgt tgccatctgc gtccttggcc tctttgccag acaagaacac
     6841 cgcatttggt ttgtacgaac catgccacgg ttctgctcca gatttgccaa agaataaggt
     6901 tgaccctatc gccactatct tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc
     6961 tgaagaaggt aaggccattg aagatgcagt taaaaaggtt ttggatgcag gtatcagaac
     7021 tggtgattta ggtggttcca acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt
     7081 taagaaaatc cttgcttaaa aagattctct ttttttatga tatttgtaca taaactttat
     7141 aaatgaaatt cataatagaa acgacacgaa attacaaaat ggaatatgtt catagggtag
     7201 acgaaactat atacgcaatc tacatacatt tatcaagaag gagaaaaagg aggatagtaa
     7261 aggaatacag gtaagcaaat tgatactaat ggctcaacgt gataaggaaa aagaattgca
     7321 ctttaacatt aatattgaca aggaggaggg caccacacaa aaagttaggt gtaacagaaa
     7381 atcatgaaac tacgattcct aatttgatat tggaggattt tctctaaaaa aaaaaaaata
     7441 caacaaataa aaaacactca atgacctgac catttgatgg agtttaagtc aataccttct
     7501 tgaagcattt cccataatgg tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc
     7561 ttacgacgta gtcgatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7621 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7681 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7741 cgtcatcacc gaaacgcgcg a
//
