LOCUS       sequence-104891-        7000 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 7000)
  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 7000)
  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.loxP
FEATURES             Location/Qualifiers
     source          1..7000
                     /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"
     protein_bind    3156..3189
                     /label=loxPsym
                     /bound_moiety="Cre recombinase"
                     /note="loxPsym has a symmetric spacer region, and loxPsym 
                     sites are capable of recombining in either orientation 
                     (Hoess et al., 1986)."
     CDS             complement(3261..3938)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             3945..3956
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3974..3990
                     /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(3998..4004,4005..4022,4023..4028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4043..4064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    4194..4227
                     /label=loxPsym
                     /bound_moiety="Cre recombinase"
                     /note="loxPsym has a symmetric spacer region, and loxPsym 
                     sites are capable of recombining in either orientation 
                     (Hoess et al., 1986)."
     primer_bind     complement(4280..4300)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4280..4298)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4319..4335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4319..4335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4343..4359
                     /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(4367..4373,4374..4391,4392..4397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4550..4567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4721..5309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4801..4820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5480..6271,6272..6340))
                     /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     6103..6122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6341..6445)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6482..6985
                     /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 ggtcgacggc agctgataac ttcgtataat gtacattata
     3181 cgaagttatg ggacgacctg cagcccccag tagagaccgc ctggctctag tagcgatcta
     3241 cactagcact atcagcgtta ttaagcaccg gtggagtgac gaccttcagc acgttcgtac
     3301 tgttcaacga tggtgtagtc ttcgttgtgg gaggtgatgt ccagtttgat gtcggttttg
     3361 taagcacccg gcagctgaac cggtttttta gccatgtagg tggttttaac ttcagcgtcg
     3421 tagtgaccac cgtctttcag tttcagacgc attttgattt cacctttcag agcaccgtct
     3481 tccgggtaca tacgttcggt ggaagcttcc caacccatgg tttttttctg cataaccgga
     3541 ccgtcggacg ggaagttggt accacgcagt ttaactttgt agatgaactc accgtcttgc
     3601 agggaggagt cctgggtaac ggtaacaaca ccaccgtctt cgaagttcat aacacgttcc
     3661 catttgaaac cttccgggaa ggacagtttc aggtagtccg ggatgtcagc cgggtgttta
     3721 acgtaagctt tggaaccgta ctggaactgc ggggacagga tgtcccaagc gaacggcagc
     3781 ggaccacctt tggtaacttt cagtttagcg gtctgggtac cttcgtacgg acgaccttca
     3841 ccttcacctt cgatttcgaa ctcgtgaccg ttaacggaac cttccatacg aactttgaaa
     3901 cgcatgaact ctttgataac gtcttcggag gaagccatct agtatttctc ctctttctct
     3961 agtatgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4021 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4081 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4141 cggggagagg cggtttgcgt attgggtctc attttgggga tcccggagca cccataactt
     4201 cgtataatgt acattatacg aagttatccc aggtcgctct agagcggccg ccaccgcggt
     4261 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     4321 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg
     4381 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt
     4441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4561 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5161 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5221 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5281 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5341 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5401 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5461 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5521 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5581 agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc
     5641 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5701 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5761 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5821 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5881 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5941 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6001 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6061 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6121 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6181 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6241 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6301 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6361 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6421 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcctttt
     6481 catcacgtgc tataaaaata attataattt aaatttttta atataaatat ataaattaaa
     6541 aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa
     6601 gactctaggg ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta
     6661 ttaatgccga attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt
     6721 ttacatttta cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat
     6781 aaatatatat gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt
     6841 tcttcattcc gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca
     6901 taaaaataaa taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc
     6961 gcgtgtaagt tacaggcaag cgatccgtcc taagaaacca
//
