LOCUS       sequence-104856-        6929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS415 backbone, contains an RFP flanked by BsaI sites with CAGT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6929)
  AUTHORS   Agmon N, Mitchell LA, Cai Y, Ikushima S, Chuang J, Zheng A, Choi WJ,
            Martin JA, Caravelli K, Stracquadanio G, Boeke JD
  TITLE     Yeast Golden Gate (yGG) for the Efficient Assembly of S. cerevisiae 
            Transcription Units.
  JOURNAL   ACS Synth Biol. 2015 Jul 17;4(7):853-9. doi: 10.1021/sb500372z. Epub
            2015 Mar 23.
  PUBMED    25756291
REFERENCE   2  (bases 1 to 6929)
  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: "10.1021/sb500372z";
            journalName: "ACS Synth Biol"; date: "2015-07-17- 17"; volume: "4"; 
            issue: "7"; pages: "853-9"
COMMENT     Sequence Label: pAV115
FEATURES             Location/Qualifiers
     source          1..6929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..45
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     145..164
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(657..1751)
                     /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(1764..2168)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2468..2923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2605..2626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2817..2836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3053..3075
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3067..3084
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3068..3084
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3094..3113
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3094..3112
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3137..3153
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3138..3154
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3238..3915)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             3922..3933
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3951..3967
                     /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(3975..3981,3982..3999,4000..4005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4020..4041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4156..4172)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4156..4172)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4209..4229)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4209..4227)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4248..4264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4248..4264)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4272..4288
                     /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(4296..4302,4303..4320,4321..4326))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4479..4496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4650..5238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4730..4749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5409..6200,6201..6269))
                     /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     6032..6051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6270..6374)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6411..6914
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt gtcacagctt
       61 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      121 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      181 tcgactacgt cgtaaggccg tttctgacag agtaaaattc ttgagggaac tttcaccatt
      241 atgggaaatg gttcaagaag gtattgactt aaactccatc aaatggtcag gtcattgagt
      301 gttttttatt tgttgtattt tttttttttt agagaaaatc ctccaatatc aaattaggaa
      361 tcgtagtttc atgattttct gttacaccta actttttgtg tggtgccctc ctccttgtca
      421 atattaatgt taaagtgcaa ttctttttcc ttatcacgtt gagccattag tatcaatttg
      481 cttacctgta ttcctttact atcctccttt ttctccttct tgataaatgt atgtagattg
      541 cgtatatagt ttcgtctacc ctatgaacat attccatttt gtaatttcgt gtcgtttcta
      601 ttatgaattt catttataaa gtttatgtac aaatatcata aaaaaagaga atctttttaa
      661 gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttcggtggt actgttggaa
      721 ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc atcttcaatg
      781 gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc agacaagata
      841 gtggcgatag ggtcaacctt attctttggc aaatctggag cagaaccgtg gcatggttcg
      901 tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga tggcaacaaa
      961 cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa catgttgctg
     1021 gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc ggcagaatca
     1081 atcaattgat gttgaacctt caatgtaggg aattcgttct tgatggtttc ctccacagtt
     1141 tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca aataggcaat
     1201 ggtggctcat gttgtagggc catgaaagcg gccattcttg tgattctttg cacttctgga
     1261 acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt tctcttacca
     1321 aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt agcaaattgt
     1381 ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg tcttaagttg
     1441 gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct aacactaccg
     1501 gtaccccatt taggaccacc cacagcacct aacaaaacgg catcaacctt cttggaggct
     1561 tccagcgcct catctggaag tgggacacct gtagcatcga tagcagcacc accaattaaa
     1621 tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt aagaacctta
     1681 atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac gatcttctta
     1741 ggggcagaca taggggcaga cattagaatg gtatatcctt gaaatatata tatatattgc
     1801 tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac ctattggaaa
     1861 aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc
     1921 atgaacgctt ctctattcta tatgaaaagc cggttccggc ctctcacctt tcctttttct
     1981 cccaattttt cagttgaaaa aggtatatgc gtcaggcgac ctctgaaatt aacaaaaaat
     2041 ttccagtcat cgaatttgat tctgtgcgat agcgcccctg tgtgttctcg ttatgttgag
     2101 gaaaaaaata atggttgcta agagattcga actcttgcat cttacgatac ctgagtattc
     2161 ccacagttaa ctgcggtcaa gatatttctt gaatcaggcg ccttagaccg ctcggccaaa
     2221 caaccaatta cttgttgaga aatagagtat aattatccta taaatataac gtttttgaac
     2281 acacatgaac aaggaagtac aggacaattg attttgaaga gaatgtggat tttgatgtaa
     2341 ttgttgggat tccattttta ataaggcaat aatattaggt atgtggatat actagaagtt
     2401 ctcctcgacc gtcgatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2461 catcaggaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     2521 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2581 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2641 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2701 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2761 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2821 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2881 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcgcgcca ttcgccattc
     2941 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     3001 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     3061 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     3121 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag
     3181 cccccagtag agaccgcctg gctctagtag cgatctacac tagcactatc agcgttatta
     3241 agcaccggtg gagtgacgac cttcagcacg ttcgtactgt tcaacgatgg tgtagtcttc
     3301 gttgtgggag gtgatgtcca gtttgatgtc ggttttgtaa gcacccggca gctgaaccgg
     3361 ttttttagcc atgtaggtgg ttttaacttc agcgtcgtag tgaccaccgt ctttcagttt
     3421 cagacgcatt ttgatttcac ctttcagagc accgtcttcc gggtacatac gttcggtgga
     3481 agcttcccaa cccatggttt ttttctgcat aaccggaccg tcggacggga agttggtacc
     3541 acgcagttta actttgtaga tgaactcacc gtcttgcagg gaggagtcct gggtaacggt
     3601 aacaacacca ccgtcttcga agttcataac acgttcccat ttgaaacctt ccgggaagga
     3661 cagtttcagg tagtccggga tgtcagccgg gtgtttaacg taagctttgg aaccgtactg
     3721 gaactgcggg gacaggatgt cccaagcgaa cggcagcgga ccacctttgg taactttcag
     3781 tttagcggtc tgggtacctt cgtacggacg accttcacct tcaccttcga tttcgaactc
     3841 gtgaccgtta acggaacctt ccatacgaac tttgaaacgc atgaactctt tgataacgtc
     3901 ttcggaggaa gccatctagt atttctcctc tttctctagt atgtgtgaaa ttgttatccg
     3961 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4021 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4141 gggtctcatt ttggggatcc actagttcta gagcggccgc caccgcggtg gagctccagc
     4201 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     4261 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca taggagccgg aagcataaag
     4321 tgtaaagcct ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg
     4381 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4441 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4501 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4561 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4621 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4681 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4741 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4801 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4861 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4921 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4981 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5041 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5101 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5161 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5221 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5281 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5341 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5401 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5461 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5521 tctggcccca gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca
     5581 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5641 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5701 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5761 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5821 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5881 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5941 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6001 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6061 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6121 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6181 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6241 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6301 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6361 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc atcacgtgct
     6421 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
     6481 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
     6541 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
     6601 ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac
     6661 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
     6721 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
     6781 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     6841 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     6901 acaggcaagc gatccgtcct aagaaacca
//
