LOCUS       sequence-453814-        7918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ymiRFP680 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7918)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7918)
  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"
COMMENT     Sequence Label: pDRF1 ymiRFP680
FEATURES             Location/Qualifiers
     source          1..7918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1808..1832)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1894..1913)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1895..1913)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2061..2248
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2116..2140)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2260..2277)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2260..2276)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2269..2291)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2712..4058
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4169..4972)
                     /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"
     promoter        complement(4973..5188)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5213..5231)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5299..5403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5404..5472,5473..6264)
                     /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     complement(5622..5641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6435..7023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6924..6943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7177..7194
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7311..7332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7347..7352,7353..7370,7371..7377)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7385..7401
                     /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)."
     primer_bind     7390..7412
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7409..7425
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7409..7425
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7455..7910)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7592..7613)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7804..7823
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc ccatggtcta
      841 gaatggctga aggttctgtt gctagacaac cagatttgtt gacttgtgat gatgaaccaa
      901 ttcatattcc aggtgctatt caaccacatg gtttgttatt agctttggct gctgatatga
      961 ccattgttgc tggttctgat aacttgcctg aattgactgg tttagctatt ggtgccttga
     1021 tcggtagatc tgctgctgat gttttcgatt ctgaaaccca taatagattg actattgctt
     1081 tggctgaacc aggtgctgct gtcggtgctc caattaccgt cggtttcacc atgcgtaagg
     1141 atgccggttt cattggttct tggcatagac atgatcaatt aattttcttg gaattggaac
     1201 caccacaacg tgacgttgcc gaaccacagg ctttcttccg tagaaccaac tctgctatta
     1261 gaagattaca agctgctgaa actttggaat ctgcttgtgc tgctgctgct caagaagttc
     1321 gtaagattac tggtttcgat agagttatga tctacagatt tgcttctgat ttctctggtg
     1381 aagttattgc tgaagataga tgtgctgaag ttgaatctaa gttgggtttg cattacccag
     1441 cttctactgt tccagctcaa gctagaagat tgtacactat taatccagtt agaattattc
     1501 cagatatcaa ctacagacca gtcccagtta ccccagattt gaacccagtt actggtagac
     1561 ctattgattt atctttcgct attttgagat ctgtttcccc atgtcatttg gaattcatga
     1621 gaaacatcgg tatgcatggt accatgtcta tttctatttt gagaggtgaa agattgtggg
     1681 gtttgattgt ttgtcatcat agaactccat actacgttga cttggatggt agacaagcct
     1741 gtaagagagt tgctgaaaga ttggctaccc aaattggtgt tatggaagaa taactcgagg
     1801 cggccgcacc cagctttctt gtacaaagtg gtgtagttaa ttcatgatgg ccgctgcagg
     1861 tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgg
     1921 atccagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag gttgtcggct
     1981 tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt ggtagatacg
     2041 ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     2101 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     2161 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     2221 tcgctcttat tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg
     2281 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     2341 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2401 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2461 acaccgcata atcggatcgt acttgttacc catcattgaa ttttgaacat ccgaacctgg
     2521 gagttttccc tgaaacagat agtatatttg aacctgtata ataatatata gtctagcgct
     2581 ttacggaaga caatgtatgt atttcggttc ctggagaaac tattgcatct attgcatagg
     2641 taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca cttcaatagc
     2701 atatctttgt taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     2761 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     2821 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     2881 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2941 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     3001 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     3061 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     3121 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     3181 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     3241 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     3301 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     3361 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     3421 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     3481 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     3541 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     3601 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt
     3661 ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact tcggaatagg
     3721 aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata
     3781 cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag
     3841 aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag
     3901 gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg
     3961 cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc
     4021 tcatccttca atgctatcat ttcctttgat attggatcga tccgatgata agctgtcaaa
     4081 catgagaatt gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     4141 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     4201 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     4261 atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc
     4321 tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca
     4381 accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa
     4441 aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg
     4501 agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt
     4561 ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa
     4621 tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac
     4681 caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct
     4741 ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta
     4801 gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa
     4861 catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag
     4921 caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc
     4981 ttcgtttcct gcatgttttt gttctgtgca gttgggttaa gaatactggg caatttcatg
     5041 tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt
     5101 cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa
     5161 aagaataaaa aaaaaatgat gaattgaaaa gctaattctt gaagacgaaa gggcctcgtg
     5221 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5281 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5401 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5461 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5521 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5581 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5641 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5701 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5761 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5821 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5881 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5941 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6001 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6061 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6121 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6181 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6241 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6301 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6361 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6421 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6481 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6541 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6601 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6661 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6721 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6781 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6841 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6901 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6961 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7021 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7081 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7141 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7201 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7261 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7321 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7381 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7441 cttaccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     7501 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     7561 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     7621 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     7681 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     7741 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     7801 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     7861 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgc
//
