LOCUS       sequence-453813-        7900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing yemiRFP670 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7900)
  TITLE     Teusink Lab yeast expression plasmids
REFERENCE   2  (bases 1 to 7900)
  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 yemiRFP670
FEATURES             Location/Qualifiers
     source          1..7900
                     /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(1790..1814)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1876..1895)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1877..1895)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2043..2230
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2098..2122)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2242..2259)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2242..2258)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2251..2273)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2694..4040
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4151..4954)
                     /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(4955..5170)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5195..5213)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5281..5385
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5386..5454,5455..6246)
                     /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(5604..5623)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6417..7005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6906..6925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7159..7176
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7293..7314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7329..7334,7335..7352,7353..7359)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7367..7383
                     /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     7372..7394
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7391..7407
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7391..7407
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7437..7892)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7574..7595)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7786..7805
                     /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 gacttgtgaa catgaagaaa
      901 ttcatttggc tggttctatt caaccacatg gtgctttgtt ggtcgtttct gaacatgacc
      961 atagagttat tcaagcctct gctaatgccg ctgaattctt aaacttgggt tctgttttag
     1021 gtgttccatt agctgaaatt gatggtgatt tgttaattaa gattttacca catttagatc
     1081 caaccgctga aggtatgcca gttgctgtta gatgtagaat cggtaatcct tccactgaat
     1141 actgtggttt gatgcataga ccaccagaag gtggtttgat tattgaattg gaaagagctg
     1201 gtccatctat tgatttatcc ggtactttgg ctccagcttt ggaaagaatt agaactgctg
     1261 gttctttaag agctttgtgt gatgacactg ttttgttatt ccaacaatgt actggttatg
     1321 atagagttat ggtttacaga ttcgatgaac aaggtcatgg tttagttttc tctgaatgtc
     1381 atgtccctgg tttggaatct tacttcggta atagataccc atcctctact gttccacaaa
     1441 tggctagaca attatacgtt agacaaagag ttagagtttt ggtcgatgtt acttaccaac
     1501 cagttccatt ggaaccaaga ttatccccat tgactggtag agatttagat atgtctggtt
     1561 gtttcttaag atctatgtct ccttgtcatt tgcaattctt gaaggatatg ggtgttagag
     1621 ctactttggc cgtttcttta gttgttggtg gtaagttgtg gggtttggtt gtttgtcacc
     1681 attacttgcc aagattcatt agattcgaat taagagctat ctgtaagaga ttggctgaaa
     1741 gaattgctac tagaattacc gctttagaat cttaactcga ggcggccgca cccagctttc
     1801 ttgtacaaag tggtgtagtt aattcatgat ggccgctgca ggtcgacctc gagggggggc
     1861 ccggtaccca attcgcccta tagtgagtcg tattacgcgc ggatccagct ttggacttct
     1921 tcgccagagg tttggtcaag tctccaatca aggttgtcgg cttgtctacc ttgccagaaa
     1981 tttacgaaaa gatggaaaag ggtcaaatcg ttggtagata cgttgttgac acttctaaat
     2041 aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg
     2101 tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact
     2161 ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt attgaccaca
     2221 cctctaccgg catgccaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2281 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2341 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     2401 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taatcggatc
     2461 gtacttgtta cccatcattg aattttgaac atccgaacct gggagttttc cctgaaacag
     2521 atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa gacaatgtat
     2581 gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg cacgtcgcat
     2641 ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt gttaacgaag
     2701 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2761 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     2821 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     2881 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     2941 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     3001 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     3061 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     3121 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     3181 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     3241 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     3301 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     3361 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     3421 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     3481 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     3541 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     3601 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     3661 ttcctatact ttctagctag agaataggaa cttcggaata ggaacttcaa agcgtttccg
     3721 aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc
     3781 gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt
     3841 gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag gtagtctagt
     3901 acctcctgtg atattatccc attccatgcg gggtatcgta tgcttccttc agcactaccc
     3961 tttagctgtt ctatatgctg ccactcctca attggattag tctcatcctt caatgctatc
     4021 atttcctttg atattggatc gatccgatga taagctgtca aacatgagaa ttgggtaata
     4081 actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata
     4141 atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc
     4201 tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa
     4261 taataatgtc agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg
     4321 cgtctccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat
     4381 ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg
     4441 caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt
     4501 ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac
     4561 cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta
     4621 ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc
     4681 tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc
     4741 cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac
     4801 ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca
     4861 agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag
     4921 tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcatgttt
     4981 ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata
     5041 tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga
     5101 ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg
     5161 atgaattgaa aagctaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     5221 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5281 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5341 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5401 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5461 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5521 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5581 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5641 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5701 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5761 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5821 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5881 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5941 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6001 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6061 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6121 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6181 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6241 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6301 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6361 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6481 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6541 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6601 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6661 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6721 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6781 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6841 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6901 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6961 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7021 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7081 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7141 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7201 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     7261 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7321 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7381 caatttcaca caggaaacag ctatgaccat gattacgcca agcttaccgc atcaggaaat
     7441 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7501 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7561 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7621 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7681 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7741 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     7801 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     7861 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
//
