LOCUS       sequence-104870-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS426 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 6634)
  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 6634)
  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: pAV126
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..31
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     131..150
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        176..396
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             397..1200
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1331..1786)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1468..1489)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1680..1699
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1916..1938
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1930..1947
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1931..1947
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1957..1976
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1957..1975
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2000..2016
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2001..2017
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2101..2778)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2785..2796
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2814..2830
                     /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(2838..2844,2845..2862,2863..2868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2883..2904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3019..3035)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3019..3035)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3072..3092)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3072..3090)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3111..3127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3111..3127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3135..3151
                     /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(3159..3165,3166..3183,3184..3189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3342..3359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3513..4101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3593..3612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4272..5063,5064..5132))
                     /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     4895..4914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5133..5237)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5264..6606
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6227..6274)
                     /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 ttaacctata aaaataggcg tatcacgagg ccctttgtca cagcttgtct gtaagcggat
       61 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
      121 cttaactatg cggcatcaga gcagattgta ctgagagtgc accataccac agcttttcaa
      181 ttcaattcat catttttttt ttattctttt ttttgatttc ggtttctttg aaattttttt
      241 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
      301 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
      361 cacagaacaa aaacctgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
      421 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
      481 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
      541 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
      601 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
      661 ttcgaagaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
      721 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
      781 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
      841 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
      901 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
      961 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     1021 aagggtgacg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     1081 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     1141 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     1201 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     1261 ttatatcagt tattacccta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     1321 ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1381 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     1441 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     1501 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     1561 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     1621 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     1681 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     1741 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg ccattcgcca
     1801 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     1861 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     1921 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
     1981 ttgggtaccg ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg
     2041 cagcccccag tagagaccgc ctggctctag tagcgatcta cactagcact atcagcgtta
     2101 ttaagcaccg gtggagtgac gaccttcagc acgttcgtac tgttcaacga tggtgtagtc
     2161 ttcgttgtgg gaggtgatgt ccagtttgat gtcggttttg taagcacccg gcagctgaac
     2221 cggtttttta gccatgtagg tggttttaac ttcagcgtcg tagtgaccac cgtctttcag
     2281 tttcagacgc attttgattt cacctttcag agcaccgtct tccgggtaca tacgttcggt
     2341 ggaagcttcc caacccatgg tttttttctg cataaccgga ccgtcggacg ggaagttggt
     2401 accacgcagt ttaactttgt agatgaactc accgtcttgc agggaggagt cctgggtaac
     2461 ggtaacaaca ccaccgtctt cgaagttcat aacacgttcc catttgaaac cttccgggaa
     2521 ggacagtttc aggtagtccg ggatgtcagc cgggtgttta acgtaagctt tggaaccgta
     2581 ctggaactgc ggggacagga tgtcccaagc gaacggcagc ggaccacctt tggtaacttt
     2641 cagtttagcg gtctgggtac cttcgtacgg acgaccttca ccttcacctt cgatttcgaa
     2701 ctcgtgaccg ttaacggaac cttccatacg aactttgaaa cgcatgaact ctttgataac
     2761 gtcttcggag gaagccatct agtatttctc ctctttctct agtatgtgtg aaattgttat
     2821 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2881 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2941 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3001 attgggtctc attttgggga tccactagtt ctagagcggc cgccaccgcg gtggagctcc
     3061 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     3121 tttcctgtgt gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata
     3181 aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca
     3241 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3301 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3361 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3421 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3481 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3541 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3601 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3661 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3721 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3781 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3841 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3901 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3961 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4021 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4081 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4141 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4201 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4261 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4321 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4381 ccatctggcc ccagtgctgc aatgataccg cgagatccac gctcaccggc tccagattta
     4441 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4501 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4561 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4621 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4681 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4741 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4801 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4861 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4921 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4981 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5041 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5101 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5161 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5221 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaacgaag catctgtgct
     5281 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     5341 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     5401 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     5461 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     5521 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     5581 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     5641 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     5701 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     5761 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     5821 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     5881 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     5941 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     6001 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     6061 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     6121 tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt
     6181 tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact
     6241 ttctagagaa taggaacttc ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc
     6301 cgaaaatgca acgcgagctg cgcacataca gctcactgtt cacgtcgcac ctatatctgc
     6361 gtgttgcctg tatatatata tacatgagaa gaacggcata gtgcgtgttt atgcttaaat
     6421 gcgtacttat atgcgtctat ttatgtagga tgaaaggtag tctagtacct cctgtgatat
     6481 tatcccattc catgcggggt atcgtatgct tccttcagca ctacccttta gctgttctat
     6541 atgctgccac tcctcaattg gattagtctc atccttcaat gctatcattt cctttgatat
     6601 tggatcatac taagaaacca ttattatcat gaca
//
