LOCUS       sequence-407643-        8453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Electra1-T2A-mCherry in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8453)
  AUTHORS   Botman D, de Groot DH, Wehrens M, van Rossum K, Yska S, van der 
            Kieft B, Albert V, Minghetti N, Goedhart J, Teusink B
  TITLE     Side-by-side systematic characterization of novel FPs in budding 
            yeast
  JOURNAL   bioRxiv 2025.06.27.661975; doi: 
            https://doi.org/10.1101/2025.06.27.661975
REFERENCE   2  (bases 1 to 8453)
  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"; journalName: "bioRxiv 
            2025.06.27.661975; doi: https://doi.org/10.1101/2025.06.27.661975"
COMMENT     Sequence Label: pDRF1-GW Electra1-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..8453
                     /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"
     CDS             1548..1601
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1617..1626
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1623..2333
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1768..1786)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(2343..2367)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2429..2448)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2430..2448)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2596..2783
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2651..2675)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2795..2812)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2795..2811)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2804..2826)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3247..4593
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4704..5507)
                     /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(5508..5723)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5748..5766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5834..5938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5939..6007,6008..6799)
                     /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(6157..6176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6970..7558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7459..7478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7712..7729
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7846..7867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7882..7887,7888..7905,7906..7912)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7920..7936
                     /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     7925..7947
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7944..7960
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7944..7960
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7990..8445)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8127..8148)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8339..8358
                     /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 atggtgtcta
      841 agggcgaaga gctgatcgag gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca
      901 acggccacca attcaaatgc acaggtgaag gagaaggcag accgtacgag ggagtgcaaa
      961 ccatgaggat caaagtcatc gagggaggac ccctgccatt tgcctttgac attcttgcca
     1021 cgtcgttcct gttcggcagc aagaccttta tcaagtaccc ggccgacatc cctgatttct
     1081 tcaaacagtc ctttcctgag ggctttactt gggaaagagt tacgagatac gaagatggtg
     1141 gagtcgtcac cgtcacgcag gacaccagcc ttgaggatgg ggggctcgtc tacaacgtca
     1201 aggtcagagg ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag accgagggtt
     1261 gggagccttt cacagagatg atgtatccag caaatggtgg tctgagagga tataccgaca
     1321 tcgcactgaa agttgatggt gatggccatc tgcacgccaa catcgtgaca acttacaggt
     1381 caaaaaagac cgtcggggac atcaagatgc ccggtgtcca tgccgttgat taccgcctgg
     1441 aaagggtcga agagagtgac aatgaaacct acgtagtgct aagagaagtg gcagttgcca
     1501 aatacagcaa ccttggtggt ggcatggacg agctgtacaa gtccggagag ggcagaggaa
     1561 gtcttctaac atgcggtgac gtggaggaga atcccggccc tggggatcca ccggtcgcca
     1621 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1681 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1741 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1801 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1861 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1921 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1981 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2041 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2101 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2161 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2221 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2281 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taaagcggcc
     2341 gcacccagct ttcttgtaca aagtggtgta gttaattcat gatggccgct gcaggtcgac
     2401 ctcgaggggg ggcccggtac ccaattcgcc ctatagtgag tcgtattacg cgcggatcca
     2461 gctttggact tcttcgccag aggtttggtc aagtctccaa tcaaggttgt cggcttgtct
     2521 accttgccag aaatttacga aaagatggaa aagggtcaaa tcgttggtag atacgttgtt
     2581 gacacttcta aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata
     2641 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
     2701 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca tgaggtcgct
     2761 cttattgacc acacctctac cggcatgcca attcactggc cgtcgtttta caacgtcgtg
     2821 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     2881 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     2941 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3001 gcataatcgg atcgtacttg ttacccatca ttgaattttg aacatccgaa cctggaagtt
     3061 ttccctgaaa cagatagtat atttgaacct gtataataat atatagtcta gcgctttacg
     3121 gaagacaatg tatgtatttc ggttcctgga gaaactattg catctattgc ataggtaatc
     3181 ttgcacgtcg catccccggt tcattttctg cgtttccatc ttgcacttca atagcatatc
     3241 tttgttaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     3301 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     3361 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     3421 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     3481 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     3541 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     3601 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     3661 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     3721 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     3781 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     3841 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     3901 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     3961 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     4021 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     4081 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     4141 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa
     4201 aagcgctctg aagttcctat actttctagc tagagaatag gaacttcgga ataggaactt
     4261 caaagcgttt ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct
     4321 cactgttcac gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa
     4381 cggcatagtg cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga
     4441 aaggtagtct agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc
     4501 ttcagcacta ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc
     4561 cttcaatgct atcatttcct ttgatattgg atcgatccga tgataagctg tcaaacatga
     4621 gaattgggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     4681 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     4741 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     4801 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     4861 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     4921 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     4981 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     5041 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     5101 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     5161 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     5221 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     5281 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     5341 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     5401 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     5461 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     5521 ttcctgcatg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     5581 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     5641 ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa
     5701 taaaaaaaaa atgatgaatt gaaaagctaa ttcttgaaga cgaaagggcc tcgtgatacg
     5761 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5821 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5881 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5941 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6001 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6061 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6121 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6181 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6241 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6301 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6361 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6421 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6481 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6541 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6601 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6661 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6721 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6781 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6841 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6901 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7081 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7621 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7681 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7741 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     7801 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     7861 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     7921 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttac
     7981 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     8041 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     8101 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     8161 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     8221 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     8281 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     8341 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     8401 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgc
//
