LOCUS       sequence-104887-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS424 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 6524)
  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 6524)
  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: pAV124
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..30
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     130..149
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        166..446
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             447..1121
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1220..1675)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1357..1378)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1569..1588
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1805..1827
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1819..1836
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1820..1836
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1846..1865
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1846..1864
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1889..1905
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1890..1906
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1990..2667)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2674..2685
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2703..2719
                     /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(2727..2733,2734..2751,2752..2757))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2772..2793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2908..2924)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2908..2924)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2961..2981)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2961..2979)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3000..3016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3000..3016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3024..3040
                     /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(3048..3054,3055..3072,3073..3078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3231..3248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3402..3990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3482..3501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4161..4952,4953..5021))
                     /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     4784..4803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5022..5126)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5153..6495
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6116..6163)
                     /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)."
ORIGIN
        1 taacctataa aaataggcgt atcacgaggc cctttgtcac agcttgtctg taagcggatg
       61 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
      121 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaacga cattactata
      181 tatataatat aggaagcatt taatagacag catcgtaata tatgtgtact ttgcagttat
      241 gacgccagat ggcagtagtg gaagatattc tttattgaaa aatagcttgt caccttacgt
      301 acaatcttga tccggagctt ttcttttttt gccgattaag aattaattcg gtcgaaaaaa
      361 gaaaaggaga gggccaagag ggagggcatt ggtgactatt gagcacgtga gtatacgtga
      421 ttaagcacac aaaggcagct tggagtatgt ctgttattaa tttcacaggt agttctggtc
      481 cattggtgaa agtttgcggc ttgcagagca cagaggccgc agaatgtgct ctagattccg
      541 atgctgactt gctgggtatt atatgtgtgc ccaatagaaa gagaacaatt gacccggtta
      601 ttgcaaggaa aatttcaagt cttgtaaaag catataaaaa tagttcaggc actccgaaat
      661 acttggttgg cgtgtttcgt aatcaaccta aggaggatgt tttggctctg gtcaatgatt
      721 acggcattga tatcgtccaa ctgcatggag atgagtcgtg gcaagaatac caagagttcc
      781 tcggtttgcc agttattaaa agactcgtat ttccaaaaga ctgcaacata ctactcagtg
      841 cagcttcaca gaaacctcat tcgtttattc ccttgtttga ttcagaagca ggtgggacag
      901 gtgaactttt ggattggaac tcgatttctg actgggttgg aaggcaagag agccccgaaa
      961 gcttacattt tatgttagct ggtggactga cgccagaaaa tgttggtgat gcgcttagat
     1021 taaatggcgt tattggtgtt gatgtaagcg gaggtgtgga gacaaatggt gtaaaagact
     1081 ctaacaaaat agcaaatttc gtcaaaaatg ctaagaaata ggttattact gagtagtatt
     1141 tatttaagta ttgtttgtgc acttgcctat gcggtgtgaa ataccgcaca gatgcgtaag
     1201 gagaaaatac cgcatcagga aattgtaaac gttaatattt tgttaaaatt cgcgttaaat
     1261 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1321 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1381 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1441 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1501 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1561 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1621 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcgc
     1681 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1741 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1801 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1861 tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc
     1921 gaattcctgc agcccccagt agagaccgcc tggctctagt agcgatctac actagcacta
     1981 tcagcgttat taagcaccgg tggagtgacg accttcagca cgttcgtact gttcaacgat
     2041 ggtgtagtct tcgttgtggg aggtgatgtc cagtttgatg tcggttttgt aagcacccgg
     2101 cagctgaacc ggttttttag ccatgtaggt ggttttaact tcagcgtcgt agtgaccacc
     2161 gtctttcagt ttcagacgca ttttgatttc acctttcaga gcaccgtctt ccgggtacat
     2221 acgttcggtg gaagcttccc aacccatggt ttttttctgc ataaccggac cgtcggacgg
     2281 gaagttggta ccacgcagtt taactttgta gatgaactca ccgtcttgca gggaggagtc
     2341 ctgggtaacg gtaacaacac caccgtcttc gaagttcata acacgttccc atttgaaacc
     2401 ttccgggaag gacagtttca ggtagtccgg gatgtcagcc gggtgtttaa cgtaagcttt
     2461 ggaaccgtac tggaactgcg gggacaggat gtcccaagcg aacggcagcg gaccaccttt
     2521 ggtaactttc agtttagcgg tctgggtacc ttcgtacgga cgaccttcac cttcaccttc
     2581 gatttcgaac tcgtgaccgt taacggaacc ttccatacga actttgaaac gcatgaactc
     2641 tttgataacg tcttcggagg aagccatcta gtatttctcc tctttctcta gtatgtgtga
     2701 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2761 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2821 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2881 ggtttgcgta ttgggtctca ttttggggat ccactagttc tagagcggcc gccaccgcgg
     2941 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     3001 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa cataggagcc
     3061 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga ggtaactcac attaattgcg
     3121 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3181 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3241 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3301 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3361 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3421 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3481 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3541 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3601 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3661 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3721 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3781 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3841 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3901 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3961 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4021 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4081 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4141 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4201 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4261 gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct
     4321 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4381 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4441 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4501 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4561 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4621 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4681 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4741 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4801 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4861 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4921 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4981 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5041 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5101 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc
     5161 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     5221 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     5281 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     5341 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     5401 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     5461 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     5521 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     5581 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     5641 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     5701 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     5761 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     5821 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     5881 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     5941 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     6001 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     6061 ttttggtttt ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt
     6121 tcctatactt tctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac
     6181 gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc
     6241 tatatctgcg tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta
     6301 tgcttaaatg cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc
     6361 ctgtgatatt atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag
     6421 ctgttctata tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc
     6481 ctttgatatt ggatcatatt aagaaaccat tattatcatg acat
//
