LOCUS       sequence-73669-e        8653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  single copy recombinase yeast expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8653)
  AUTHORS   Ansari A, Cheng TH, Gartenberg MR
  TITLE     Isolation of selected chromatin fragments from yeast by 
            site-specific recombination in vivo.
  JOURNAL   Methods. 1999 Feb;17(2):104-11.
  PUBMED    10075889
REFERENCE   2  (bases 1 to 8653)
  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: "Methods."; 
            date: "1999-02"; volume: "17(2)"; pages: "104-11"
COMMENT     Sequence Label: pRS415-recR
FEATURES             Location/Qualifiers
     source          1..8653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2611..2632)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2823..2842
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3059..3081
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3073..3090
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3074..3090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3100..3119
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3100..3118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3143..3159
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3144..3160
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3194..3210)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3194..3210)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        3228..3892
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3444..3561
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3856..3879
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(5879..5899)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5879..5897)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5918..5934)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5918..5934)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5931..5953)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5942..5958
                     /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)."
     promoter        complement(join(5966..5972,5973..5990,5991..5996))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6149..6166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6320..6908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6400..6419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7079..7870,7871..7939))
                     /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     7702..7721
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7940..8044)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8081..8584
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8626..8644
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
      241 accattatgg gaaatggttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
      301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
      361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
      421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
      481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
      541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
      601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
      661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
      721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
      781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
      841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
      901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
      961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     1141 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
     1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     1501 ctaccggtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
     1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     2401 gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3001 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3061 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     3121 gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc
     3181 ctgcagcccg ggggatccac tagttctaga gcggccgcta attcaatttt caaaaattct
     3241 tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc
     3301 atatacatat ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa
     3361 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa
     3421 ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac agccctccga
     3481 aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt
     3541 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg
     3601 aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa
     3661 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc
     3721 gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt
     3781 aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat
     3841 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatc
     3901 caattatgca gctgaccaag gatactgaaa tttcaacaat taatagacaa atgtcagact
     3961 tcagcgagct tagtcaaatc cttcccttac accagatctc taaaataaaa gacattttgg
     4021 agaatgagaa ccctttacct aaggaaaaac tagcctccca tctgacgatg ataattttga
     4081 tggccaattt ggcgagtcag aaaagaaagg acgttcctgt caagcgctcc actttcttga
     4141 aataccagag gtctatttca aaaacattgc aatacgattc ttccactaaa acggtatcct
     4201 ttgaatacca tttgaaagac ccaagtaagc taataaaagg actcgaagat gttgtatcgc
     4261 cctatagatt tgttgtgggg gtacacgaaa agcctgatga tgtaatgagt catttatctg
     4321 ctgtacatat gagaaaggaa gcaggtcgga aaagagattt gggtaacaag attaatgatg
     4381 aaatcactaa aatcgctgaa acacaggaaa ctatttgggg gtttgtcggg aagactatgg
     4441 acttaataga agctagaact actcggccta caacaaaagc cgcgtacaat cttttgcttc
     4501 aggcgacttt tatgaattgt tgtagagcag acgacttaaa gaataccgat atcaagacat
     4561 ttgaagttat tccagacaag caccttggcc gcatgctacg tgcttttgtc ccagagacta
     4621 agactggtac tcgttttgtc tatttcttcc cttgtaaagg acgatgcgac ccgctgttgg
     4681 cactagattc atacctccaa tggacagatc ctattcccaa aactcgaact actgatgagg
     4741 atgcacgcta tgattaccag cttctcagga acagtctgtt aggctcatat gacgggttta
     4801 tctctaaaca atcggatgag agcatcttta aaatacccaa tggaccgaaa gcgcatttag
     4861 gcaggcatgt gacagcttca tatctttcaa ataatgagat ggataaagaa gccacgctct
     4921 atggtaattg gtctgccgcg cgggaagaag gtgttagccg tgtggctaag gctcgataca
     4981 tgcacacgat agaaaagagt ccgccttctt atttatttgc gtttctatcg gggttttata
     5041 atatcacagc cgaaagggcg tgtgagttgg ttgatccaaa tagcaaccct tgtgaacaag
     5101 acaagaatat tcctatgata tcagatatcg agacattgat ggctcggtat ggaaagaatg
     5161 cagaaataat accaatggat gtattggttt ttttgagcag ctatgccagg ttcaagaata
     5221 acgaaggtaa agagtataag ttacaagcaa gaagttcccg aggcgtccca gattttccgg
     5281 ataatgggag aactgcactt tataacgcgt taacagcggc acatgtgaag cgccgaaaga
     5341 ttagcattgt cgtcggacga tccatcgaca cctcctgaca cctcctgaca agattaagcc
     5401 ttcacatcct ccgaacacac caccggactc ggactctatt tcaagcattg acgtaacttg
     5461 ctcaaataca ggctcgttgg tgctctctca agaattatct ttcaacagat gccctgatat
     5521 atcaggagac tcgtcttaaa taggctggat aatgatatac acgtaatgta atgtaatgta
     5581 atgtcatgta agatatacga tttatgattc aatgttttaa gattctacac tagatttgag
     5641 ctactggacg ggaattgtgg aggggtattg tggaatggag tgttgtgttg tggaacgggg
     5701 ttaattgtaa tgataatgat aatgataacg gtgggttata gagcattgca tctgtcgagt
     5761 tacgatgata gagttgtaag ggcggcagcg ggtatggcct aaataattta gtttagctta
     5821 atataattta gtttttgtcg aagcggccgc caccgcggtg gagctccagc ttttgttccc
     5881 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5941 attgttatcc gctcacaatt ccacacaaca taggagccgg aagcataaag tgtaaagcct
     6001 ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc
     6061 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6121 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6181 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6241 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6301 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6361 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6421 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6481 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6541 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6601 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6661 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6721 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6781 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6841 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6961 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7021 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7081 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7141 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7201 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7261 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7321 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7381 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7441 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7501 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7561 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7621 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7681 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7741 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7801 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7861 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7921 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7981 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8041 cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc atcacgtgct ataaaaataa
     8101 ttataattta aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa
     8161 ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac
     8221 aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc
     8281 ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa
     8341 tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc
     8401 tttttgttga aattttttaa acctttgttt attttttttt cttcattccg taactcttct
     8461 accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag
     8521 taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc
     8581 gatccgtcct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     8641 aggccctttc gtc
//
