LOCUS       sequence-104894-        5182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS404 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 5182)
  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 5182)
  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: pAV104
FEATURES             Location/Qualifiers
     source          1..5182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..40
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     140..159
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        176..456
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             457..1131
                     /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(1230..1685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1367..1388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1579..1598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1815..1837
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1829..1846
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1830..1846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1856..1875
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1856..1874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1899..1915
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1900..1916
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2000..2677)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2684..2695
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2713..2729
                     /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(2737..2743,2744..2761,2762..2767))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2782..2803
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2918..2934)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2918..2934)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2971..2991)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2971..2989)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3010..3026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3010..3026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3034..3050
                     /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(3058..3064,3065..3082,3083..3088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3241..3258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3412..4000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3492..3511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4171..4962,4963..5031))
                     /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     4794..4813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5032..5136)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atcatgacat taacctataa aaataggcgt atcacgaggc cctttgtcac agcttgtctg
       61 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
      121 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaacga
      181 cattactata tatataatat aggaagcatt taatagacag catcgtaata tatgtgtact
      241 ttgcagttat gacgccagat ggcagtagtg gaagatattc tttattgaaa aatagcttgt
      301 caccttacgt acaatcttga tccggagctt ttcttttttt gccgattaag aattaattcg
      361 gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt ggtgactatt gagcacgtga
      421 gtatacgtga ttaagcacac aaaggcagct tggagtatgt ctgttattaa tttcacaggt
      481 agttctggtc cattggtgaa agtttgcggc ttgcagagca cagaggccgc agaatgtgct
      541 ctagattccg atgctgactt gctgggtatt atatgtgtgc ccaatagaaa gagaacaatt
      601 gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag catataaaaa tagttcaggc
      661 actccgaaat acttggttgg cgtgtttcgt aatcaaccta aggaggatgt tttggctctg
      721 gtcaatgatt acggcattga tatcgtccaa ctgcatggag atgagtcgtg gcaagaatac
      781 caagagttcc tcggtttgcc agttattaaa agactcgtat ttccaaaaga ctgcaacata
      841 ctactcagtg cagcttcaca gaaacctcat tcgtttattc ccttgtttga ttcagaagca
      901 ggtgggacag gtgaactttt ggattggaac tcgatttctg actgggttgg aaggcaagag
      961 agccccgaaa gcttacattt tatgttagct ggtggactga cgccagaaaa tgttggtgat
     1021 gcgcttagat taaatggcgt tattggtgtt gatgtaagcg gaggtgtgga gacaaatggt
     1081 gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg ctaagaaata ggttattact
     1141 gagtagtatt tatttaagta ttgtttgtgc acttgcctat gcggtgtgaa ataccgcaca
     1201 gatgcgtaag gagaaaatac cgcatcagga aattgtaaac gttaatattt tgttaaaatt
     1261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1321 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1441 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     1621 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1681 cgcgtcgcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     1741 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     1801 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata
     1861 cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgataa
     1921 gcttgatatc gaattcctgc agcccccagt agagaccgcc tggctctagt agcgatctac
     1981 actagcacta tcagcgttat taagcaccgg tggagtgacg accttcagca cgttcgtact
     2041 gttcaacgat ggtgtagtct tcgttgtggg aggtgatgtc cagtttgatg tcggttttgt
     2101 aagcacccgg cagctgaacc ggttttttag ccatgtaggt ggttttaact tcagcgtcgt
     2161 agtgaccacc gtctttcagt ttcagacgca ttttgatttc acctttcaga gcaccgtctt
     2221 ccgggtacat acgttcggtg gaagcttccc aacccatggt ttttttctgc ataaccggac
     2281 cgtcggacgg gaagttggta ccacgcagtt taactttgta gatgaactca ccgtcttgca
     2341 gggaggagtc ctgggtaacg gtaacaacac caccgtcttc gaagttcata acacgttccc
     2401 atttgaaacc ttccgggaag gacagtttca ggtagtccgg gatgtcagcc gggtgtttaa
     2461 cgtaagcttt ggaaccgtac tggaactgcg gggacaggat gtcccaagcg aacggcagcg
     2521 gaccaccttt ggtaactttc agtttagcgg tctgggtacc ttcgtacgga cgaccttcac
     2581 cttcaccttc gatttcgaac tcgtgaccgt taacggaacc ttccatacga actttgaaac
     2641 gcatgaactc tttgataacg tcttcggagg aagccatcta gtatttctcc tctttctcta
     2701 gtatgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2761 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2821 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2881 ggggagaggc ggtttgcgta ttgggtctca ttttggggat ccactagttc tagagcggcc
     2941 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     3001 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3061 cataggagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga ggtaactcac
     3121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3181 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3241 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3301 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3361 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3421 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3481 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3541 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3601 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3661 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3721 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3781 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3841 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3901 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3961 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4021 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4081 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4141 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4201 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4261 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg
     4321 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4381 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4441 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4501 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4561 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4621 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4681 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4741 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4801 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4861 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4921 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4981 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5041 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5101 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5161 tgacgtctaa gaaaccatta tt
//
