LOCUS       sequence-104851-        5878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS413 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 5878)
  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 5878)
  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: pAV113
FEATURES             Location/Qualifiers
     source          1..5878
                     /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"
     promoter        311..497
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             498..1157
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1417..1872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1554..1575)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1766..1785
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2002..2024
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2016..2033
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2017..2033
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2043..2062
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2043..2061
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2086..2102
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2087..2103
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2187..2864)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2871..2882
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2900..2916
                     /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(2924..2930,2931..2948,2949..2954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2969..2990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3105..3121)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3105..3121)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3158..3178)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3158..3176)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3197..3213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3197..3213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3221..3237
                     /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(3245..3251,3252..3269,3270..3275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3428..3445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3599..4187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3679..3698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4358..5149,5150..5218))
                     /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     4981..5000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5219..5323)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5360..5863
                     /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 cgtgttggcg
      121 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      181 aattcccgtt ttaagagctt ggtgagcgct aggagtcact gccaggtatc gtttgaacac
      241 ggcattagtc agggaagtca taacacagtc ctttcccgca attttctttt tctattactc
      301 ttggcctcct ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt
      361 ttttccccta gcggatgact cttttttttt cttagcgatt ggcattatca cataatgaat
      421 tatacattat ataaagtaat gtgatttctt cgaagaatat actaaaaaat gagcaggcaa
      481 gataaacgaa ggcaaagatg acagagcaga aagccctagt aaagcgtatt acaaatgaaa
      541 ccaagattca gattgcgatc tctttaaagg gtggtcccct agcgatagag cactcgatct
      601 tcccagaaaa agaggcagaa gcagtagcag aacaggccac acaatcgcaa gtgattaacg
      661 tccacacagg tatagggttt ctggaccata tgatacatgc tctggccaag cattccggct
      721 ggtcgctaat cgttgagtgc attggtgact tacacataga cgaccatcac accactgaag
      781 actgcgggat tgctctcggt caagctttta aagaggccct actggcgcgt ggagtaaaaa
      841 ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg gtagatcttt
      901 cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta ggagatctct
      961 cttgcgagat gatcccgcat tttcttgaaa gctttgcaga ggctagcaga attaccctcc
     1021 acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg ttcaaggctc
     1081 ttgcggttgc cataagagaa gccacctcgc ccaatggtac caacgatgtt ccctccacca
     1141 aaggtgttct tatgtagtga caccgattat ttaaagctgc agcatacgat atatatacat
     1201 gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta tgatactgaa
     1261 gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg ctttcctttt
     1321 ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg gtgtgaaata
     1381 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt
     1441 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1501 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1561 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1621 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1681 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1741 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1801 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1861 tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1921 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     1981 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     2041 cgtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta
     2101 tcgataagct tgatatcgaa ttcctgcagc ccccagtaga gaccgcctgg ctctagtagc
     2161 gatctacact agcactatca gcgttattaa gcaccggtgg agtgacgacc ttcagcacgt
     2221 tcgtactgtt caacgatggt gtagtcttcg ttgtgggagg tgatgtccag tttgatgtcg
     2281 gttttgtaag cacccggcag ctgaaccggt tttttagcca tgtaggtggt tttaacttca
     2341 gcgtcgtagt gaccaccgtc tttcagtttc agacgcattt tgatttcacc tttcagagca
     2401 ccgtcttccg ggtacatacg ttcggtggaa gcttcccaac ccatggtttt tttctgcata
     2461 accggaccgt cggacgggaa gttggtacca cgcagtttaa ctttgtagat gaactcaccg
     2521 tcttgcaggg aggagtcctg ggtaacggta acaacaccac cgtcttcgaa gttcataaca
     2581 cgttcccatt tgaaaccttc cgggaaggac agtttcaggt agtccgggat gtcagccggg
     2641 tgtttaacgt aagctttgga accgtactgg aactgcgggg acaggatgtc ccaagcgaac
     2701 ggcagcggac cacctttggt aactttcagt ttagcggtct gggtaccttc gtacggacga
     2761 ccttcacctt caccttcgat ttcgaactcg tgaccgttaa cggaaccttc catacgaact
     2821 ttgaaacgca tgaactcttt gataacgtct tcggaggaag ccatctagta tttctcctct
     2881 ttctctagta tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2941 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3001 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3061 aacgcgcggg gagaggcggt ttgcgtattg ggtctcattt tggggatcca ctagttctag
     3121 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     3181 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3241 cacacaacat aggagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt
     3301 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3781 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4021 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4081 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4141 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4201 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4261 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4321 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4381 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4441 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4501 atccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4561 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4621 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4681 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4741 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4801 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4861 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4921 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4981 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5041 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5101 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5161 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5221 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5281 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5341 tgccacctgg gtccttttca tcacgtgcta taaaaataat tataatttaa attttttaat
     5401 ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg
     5461 ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc
     5521 tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca
     5581 cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat
     5641 ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa
     5701 cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta
     5761 aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa attattccat
     5821 cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atccgtccta agaaacca
//
