LOCUS       sequence-326050-        8392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET fusion between ymScarletI and ymNeongreen for use in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8392)
  TITLE     Teusink unpublished plasmids
REFERENCE   2  (bases 1 to 8392)
  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: ymScarletI-ymNeongreen pDRF1-GW
FEATURES             Location/Qualifiers
     source          1..8392
                     /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"
     regulatory      1557..1566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1818..1827
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2282..2306)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2368..2387)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2369..2387)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2535..2722
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2590..2614)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2734..2751)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2734..2750)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2743..2765)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3186..4532
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4643..5446)
                     /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(5447..5662)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5687..5705)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5773..5877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5878..5946,5947..6738)
                     /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(6096..6115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6909..7497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7398..7417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7651..7668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7785..7806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7821..7826,7827..7844,7845..7851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7859..7875
                     /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     7864..7886
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7883..7899
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7883..7899
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7929..8384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8066..8087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8278..8297
                     /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 atggttagta
      841 aaggtgaagc tgttataaaa gaattcatga ggtttaaagt tcatatggaa ggttcaatga
      901 atggtcatga atttgaaatt gaaggtgaag gtgaaggtag accatatgaa ggtacacaaa
      961 ctgctaaatt gaaagttact aaaggtggtc cattaccatt ttcttgggat attttgtctc
     1021 cacaattcat gtatggttct agagctttta ttaagcatcc agctgatatt ccagattatt
     1081 ataaacaatc ttttcctgaa ggttttaaat gggaaagagt tatgaatttc gaagatggtg
     1141 gtgctgttac tgttactcaa gatacttctt tggaagatgg tactttaatc tataaagtta
     1201 aattgagagg tactaatttt ccaccagatg gtccagttat gcaaaagaaa actatgggtt
     1261 gggaagcatc tactgaaaga ttgtatccag aagatggtgt tttgaaagga gacattaaga
     1321 tggctttgag attgaaagat ggtggtcggt acctggccga cttcaagacc acctataaag
     1381 ctaaaaaacc agttcaaatg ccaggtgcat ataatgttga tagaaagtta gatataacgt
     1441 cgcataacga ggactatact gtagttgaac aatatgaacg tagtgaaggt agacatagta
     1501 ccggaggaat ggatgaattg tataaatccg gactcagatc ttcggatcca ccggtcgcca
     1561 ccatggtctc taagggtgaa gaagataaca tggcttcttt gccagccacc cacgagttgc
     1621 acatcttcgg ttccattaac ggtgtcgact tcgatatggt cggtcaaggt accggtaacc
     1681 caaacgacgg ttacgaagaa ttgaacttga agtctaccaa gggcgacttg cagttctccc
     1741 cttggatttt ggttccacac atcggttacg gtttccacca atacttgcca tacccagacg
     1801 gtatgtcccc attccaagcc gccatggttg acggttctgg ttaccaagtc cacagaacca
     1861 tgcaattcga agacggtgct tccttgaccg tgaactacag atacacctac gaaggttctc
     1921 acatcaaggg cgaagcccaa gttaagggta ccggtttccc tgctgacggt ccagtcatga
     1981 ctaactcctt gaccgctgct gactggtgtc gttccaagaa gacctaccca aacgataaga
     2041 ccatcatttc cactttcaag tggtcctaca ccaccggtaa cggcaagaga taccgctcta
     2101 ccgctcgtac cacctacacc ttcgccaagc caatggctgc caactacttg aagaaccagc
     2161 ctatgtacgt cttccgtaag accgagctga agcattccaa gactgaattg aacttcaagg
     2221 aatggcaaaa ggctttcacc gacgtcatgg gtatggacga attgtacaag taagcggccg
     2281 cacccagctt tcttgtacaa agtggtgtag ttaattcatg atggccgctg caggtcgacc
     2341 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc gcggatccag
     2401 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     2461 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     2521 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2581 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga naattcttat
     2641 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     2701 ttattgacca cacctctacc ggcatgccaa ttcactggcc gtcgttttac aacgtcgtga
     2761 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2821 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2881 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2941 cataatcgga tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt
     3001 tccctgaaac agatagtata tttgaacctg tataataata tatagtctag cgctttacgg
     3061 aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct
     3121 tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct
     3181 ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa
     3241 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc
     3301 tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc
     3361 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag
     3421 agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg
     3481 agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta
     3541 taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac
     3601 tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat
     3661 tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt atattctata
     3721 ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg
     3781 gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt
     3841 ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt
     3901 ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt
     3961 caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa
     4021 agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta
     4081 cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa
     4141 agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa taggaacttc
     4201 aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc
     4261 actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac
     4321 ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa
     4381 aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct
     4441 tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc
     4501 ttcaatgcta tcatttcctt tgatattgga tcgatccgat gataagctgt caaacatgag
     4561 aattgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg
     4621 catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc
     4681 agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc
     4741 ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg gttctatact
     4801 gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat
     4861 cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt
     4921 tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct
     4981 tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg
     5041 cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg
     5101 cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt
     5161 cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg
     5221 gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac
     5281 aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca
     5341 atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag
     5401 gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt
     5461 tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     5521 caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     5581 tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa aaaaaagaat
     5641 aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac gaaagggcct cgtgatacgc
     5701 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5761 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5821 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5881 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5941 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6001 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6061 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6121 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6421 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7681 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7741 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7801 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7861 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttacc
     7921 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     7981 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     8041 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     8101 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     8161 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     8221 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     8281 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     8341 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gc
//
