LOCUS       sequence-104862-        7759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS425 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 7759)
  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 7759)
  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: pAV125
FEATURES             Location/Qualifiers
     source          1..7759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(627..1721)
                     /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"
     promoter        complement(1734..2138)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2438..2893)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2575..2596)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2787..2806
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3023..3045
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3037..3054
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3038..3054
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3064..3083
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3064..3082
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3107..3123
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3108..3124
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3208..3885)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             3892..3903
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3921..3937
                     /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(3945..3951,3952..3969,3970..3975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3990..4011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4128..4144)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4128..4144)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4181..4201)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4181..4199)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4220..4236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4220..4236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4244..4260
                     /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(4268..4274,4275..4292,4293..4298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4451..4468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4622..5210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4702..4721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5381..6172,6173..6241))
                     /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     6004..6023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6242..6346)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6373..7715
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7336..7383)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     join(7756..7759,1..15)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggcgtatcac gaggcccttt gtcacagctt gtctgtaagc ggatgccggg agcagacaag
       61 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
      121 cagagcagat tgtactgaga gtgcaccata tcgactacgt cgtaaggccg tttctgacag
      181 agtaaaattc ttgagggaac tttcaccatt atgggaaatg cttcaagaag gtattgactt
      241 aaactccatc aaatggtcag gtcattgagt gttttttatt tgttgtattt tttttttttt
      301 agagaaaatc ctccaatatc aaattaggaa tcgtagtttc atgattttct gttacaccta
      361 actttttgtg tggtgccctc ctccttgtca atattaatgt taaagtgcaa ttctttttcc
      421 ttatcacgtt gagccattag tatcaatttg cttacctgta ttcctttact atcctccttt
      481 ttctccttct tgataaatgt atgtagattg cgtatatagt ttcgtctacc ctatgaacat
      541 attccatttt gtaatttcgt gtcgtttcta ttatgaattt catttataaa gtttatgtac
      601 aaatatcata aaaaaagaga atctttttaa gcaaggattt tcttaacttc ttcggcgaca
      661 gcatcaccga cttcggtggt actgttggaa ccacctaaat caccagttct gatacctgca
      721 tccaaaacct ttttaactgc atcttcaatg gccttacctt cttcaggcaa gttcaatgac
      781 aatttcaaca tcattgcagc agacaagata gtggcgatag ggtcaacctt attctttggc
      841 aaatctggag cagaaccgtg gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc
      901 aaagaggcca aggacgcaga tggcaacaaa cccaaggaac ctgggataac ggaggcttca
      961 tcggagatga tatcaccaaa catgttgctg gtgattataa taccatttag gtgggttggg
     1021 ttcttaacta ggatcatggc ggcagaatca atcaattgat gttgaacctt caatgtaggg
     1081 aattcgttct tgatggtttc ctccacagtt tttctccata atcttgaaga ggccaaaaga
     1141 ttagctttat ccaaggacca aataggcaat ggtggctcat gttgtagggc catgaaagcg
     1201 gccattcttg tgattctttg cacttctgga acggtgtatt gttcactatc ccaagcgaca
     1261 ccatcaccat cgtcttcctt tctcttacca aagtaaatac ctcccactaa ttctctgaca
     1321 acaacgaagt cagtaccttt agcaaattgt ggcttgattg gagataagtc taaaagagag
     1381 tcggatgcaa agttacatgg tcttaagttg gcgtacaatt gaagttcttt acggattttt
     1441 agtaaacctt gttcaggtct aacactaccg gtaccccatt taggaccagc cacagcacct
     1501 aacaaaacgg catcaacctt cttggaggct tccagcgcct catctggaag tgggacacct
     1561 gtagcatcga tagcagcacc accaattaaa tgattttcga aatcgaactt gacattggaa
     1621 cgaacatcag aaatagcttt aagaacctta atggcttcgg ctgtgatttc ttgaccaacg
     1681 tggtcacctg gcaaaacgac gatcttctta ggggcagaca taggggcaga cattagaatg
     1741 gtatatcctt gaaatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa
     1801 gtaagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa
     1861 cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc
     1921 cggttccggc ctctcacctt tcctttttct cccaattttt cagttgaaaa aggtatatgc
     1981 gtcaggcgac ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat
     2041 agcgcccctg tgtgttctcg ttatgttgag gaaaaaaata atggttgcta agagattcga
     2101 actcttgcat cttacgatac ctgagtattc ccacagttaa ctgcggtcaa gatatttctt
     2161 gaatcaggcg ccttagaccg ctcggccaaa caaccaatta cttgttgaga aatagagtat
     2221 aattatccta taaatataac gtttttgaac acacatgaac aaggaagtac aggacaattg
     2281 attttgaaga gaatgtggat tttgatgtaa ttgttgggat tccattttta ataaggcaat
     2341 aatattaggt atgtggatat actagaagtt ctcctcgacc gtcgatatgc ggtgtgaaat
     2401 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt taatattttg
     2461 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     2521 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     2581 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     2641 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     2701 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2761 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2821 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2881 ctacagggcg cgtcgcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2941 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     3001 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     3061 gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga ggtcgacggt
     3121 atcgataagc ttgatatcga attcctgcag cccccagtag agaccgcctg gctctagtag
     3181 cgatctacac tagcactatc agcgttatta agcaccggtg gagtgacgac cttcagcacg
     3241 ttcgtactgt tcaacgatgg tgtagtcttc gttgtgggag gtgatgtcca gtttgatgtc
     3301 ggttttgtaa gcacccggca gctgaaccgg ttttttagcc atgtaggtgg ttttaacttc
     3361 agcgtcgtag tgaccaccgt ctttcagttt cagacgcatt ttgatttcac ctttcagagc
     3421 accgtcttcc gggtacatac gttcggtgga agcttcccaa cccatggttt ttttctgcat
     3481 aaccggaccg tcggacggga agttggtacc acgcagttta actttgtaga tgaactcacc
     3541 gtcttgcagg gaggagtcct gggtaacggt aacaacacca ccgtcttcga agttcataac
     3601 acgttcccat ttgaaacctt ccgggaagga cagtttcagg tagtccggga tgtcagccgg
     3661 gtgtttaacg taagctttgg aaccgtactg gaactgcggg gacaggatgt cccaagcgaa
     3721 cggcagcgga ccacctttgg taactttcag tttagcggtc tgggtacctt cgtacggacg
     3781 accttcacct tcaccttcga tttcgaactc gtgaccgtta acggaacctt ccatacgaac
     3841 tttgaaacgc atgaactctt tgataacgtc ttcggaggaa gccatctagt atttctcctc
     3901 tttctctagt atgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3961 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4021 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4081 caacgcgcgg ggagaggcgg tttgcgtatt gggtctcatt ttggggggat ccactagttc
     4141 tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
     4201 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4261 ttccacacaa cataggagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4321 ggtaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4381 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4441 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4501 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5041 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5281 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5341 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5401 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5461 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5521 gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5581 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5641 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5701 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5761 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5821 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5881 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5941 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6001 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6061 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6121 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6181 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6241 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6301 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6361 aagtgccacc tgaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     6421 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     6481 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     6541 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     6601 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     6661 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     6721 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     6781 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     6841 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     6901 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     6961 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     7021 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     7081 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     7141 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     7201 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     7261 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt
     7321 tttcaaaagc gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac
     7381 ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag
     7441 ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag
     7501 aacggcatag tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt tatgtaggat
     7561 gaaaggtagt ctagtacctc ctgtgatatt atcccattcc atgcggggta tcgtatgctt
     7621 ccttcagcac taccctttag ctgttctata tgctgccact cctcaattgg attagtctca
     7681 tccttcaatg ctatcatttc ctttgatatt ggatcatact aagaaaccat tattatcatg
     7741 acattaacct ataaaaata
//
