LOCUS       sequence-375302-        6364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression vector for bacteria and mammalian cells producing 
            mRuby3-SYFP2 fusion protein for FRET analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6364)
  AUTHORS   Gadella TWJ Jr, van Weeren L, Stouthamer J, Hink MA, Wolters AHG, 
            Giepmans BNG, Aumonier S, Dupuy J, Royant A
  TITLE     mScarlet3: a brilliant and fast-maturing red fluorescent protein.
  JOURNAL   Nat Methods. 2023 Apr;20(4):541-545. doi: 
            10.1038/s41592-023-01809-y. Epub 2023 Mar 27.
  PUBMED    36973546
REFERENCE   2  (bases 1 to 6364)
  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: 
            "10.1038/s41592-023-01809-y"; journalName: "Nat Methods"; date: 
            "2023-04"; volume: "20"; issue: "4"; pages: "541-545"
COMMENT     Sequence Label: pDx_mRuby3-SYFP2
FEATURES             Location/Qualifiers
     source          1..6364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     236..258
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     320..339
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        381..499
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             574..591
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             601..618
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      637..646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1348..1350,1351..1353,1354..2064)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1393..1414)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1654..1673)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2001..2022
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2154..2173)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2219..2274
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2273..2292)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2362..2409
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2422..3127
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2510..2882
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2539..3127)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3136..3169)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3337..3924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3416..3435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3937..4752)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4660..4679
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_recomb     4995..5822
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4995..5119
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5157..5762)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5890..6193
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6195..6364,1..33)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(6348..6364,1..4)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cgtgtacggt gggaggtcta tataagcaga cgtcgtttag tgaaccgtca gatcactaga
       61 tgctttattg cggtagttta tcacagttaa attgctaacg ccagtctcga acttaacgtg
      121 cagaagttgg tcgtgaggca ctgggcaggt aagtatcggg ccctttgtgc ggggggagcg
      181 gctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc
      241 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
      301 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
      361 gaattggatc ggaccgaaat caccacaatt cagcaaattg tgaacatcat cacgttcatc
      421 tttccctggt tgccaaaggc ccattttcct gtcagtaacg agaaggtcgc gaattcaggc
      481 gctttttaga ctggtcgtag ggagaccaca acggtttccc tctagaaata attttgttta
      541 actataagaa ggagatatac catgcggggt tctcatcatc accaccatca cagcagcggc
      601 ctggtgccgc gcggcagctc gctagcgcta ccggtcgcca ccatggtgtc taagggcgaa
      661 gagctgatca aggaaaatat gcgtatgaag gtggtcatgg aaggttcggt caacggccac
      721 caattcaaat gcacaggtga aggagaaggc cgcccgtacg agggagtgca aaccatgagg
      781 atcaaagtca tcgagggagg acccctgcca tttgcctttg acattcttgc cacgtcgttc
      841 atgtatggca gccgtacttt tatcaagtac ccggcagaca ttcctgattt ctttaaacag
      901 tcctttcctg aaggttttac ttgggaaaga gttacgagat acgaagatgg tggagtcgtc
      961 accgtcaccc aggacaccag ccttgaggat ggcgaactcg tttacaacgt caaagtcaga
     1021 ggggtaaact ttccctccaa tggtcccgtg atgcagaaga agaccaaggg ttgggagcct
     1081 aatacagaga tgatgtatcc agcagatggt ggtctgaggg gatacactga tatcgcactg
     1141 aaagttgatg gtggtggcca tctgcattgc aatttcgtaa caacttacag gtcaaaaaag
     1201 accgtcggga acatcaagat gcccggtgtg catgccgttg atcaccgcct ggaaaggatt
     1261 gaggaaagtg acaatgaaac ctacgtagta caacgcgaag tggcagttgc caaatacagc
     1321 aaccttggta tggacgagct gtacaagatg gtgagcaagg gcgaggagct gttcaccggg
     1381 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1441 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     1501 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc
     1561 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1621 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1681 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1741 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1801 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1861 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1921 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
     1981 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2041 ctcggcatgg acgagctcta caagtaagtg attaacctca ggtgcaggct gcctatcaga
     2101 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat cgatcttttt
     2161 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     2221 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     2281 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     2341 gcaacatatg cccatatgta actagcataa ccccttgggg cctctaaacg ggtcttgagg
     2401 ggttttttgc tgaaagcatg cggaggaaat tctccttgaa gtttccctgg tgttcaaagt
     2461 aaaggagttt gcaccagacg cacctctgtt cactggtccg gcgtattaaa acacgataca
     2521 ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta cagacaattg
     2581 ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg ttagagcgaa
     2641 aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct caatagattt
     2701 gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga tggctggact
     2761 atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca gcaatctagc
     2821 tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt
     2881 atgcgctttt gtatttcttt catcactgtc gttagtatac aattgactcg acgtaaacac
     2941 gttaaataga gcttggacat atttaacatc gggcgtgtta gctttattag gccgattatc
     3001 gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg ccatagccac
     3061 acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag ttgagctttt
     3121 tggaattgcg atcgcataac ttcgtatagc atacattata cgaagttata agctcggaac
     3181 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3241 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gccccctgac
     3361 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3421 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3481 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3541 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3601 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3661 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3721 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3781 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3841 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3901 acgcgcagaa aaaaaggatc tcaagaagat cctttgttag aaaaactcat cgagcatcaa
     3961 gtgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     4021 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     4081 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     4141 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     4201 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     4261 actcgcacca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     4321 atcgccgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     4381 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     4441 tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     4501 gatggtcgga agaggcataa attccgtcag ccagtttagc ctgaccatct catctgtaac
     4561 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     4621 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     4681 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     4741 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4801 catgtccgcg cgtttcctgc atcttttaat caaatcccaa gatgtgtata aacgcgccgg
     4861 tatgtatagg aagaggttta tactaaactg ttacattgca aacgtggttt cgtgtgccaa
     4921 gtgtgaaaac cgatgtttaa tcaaggctct gacgcatttc tacaaccacg actccaagtg
     4981 tgtgggtgaa gtcatgcatc ttttaatcaa atcccaagat gtgtataaac caccaaactg
     5041 ccaaaaaatg aaaactgtcg acaagctctg tccgtttgct ggcaactgca agggtctcaa
     5101 tcctatttgt aattattgaa taataaaaca attataaatg tcaaatttgt tttttattaa
     5161 cgatacaaac caaacgcaac aagaacattt gtagtattat ctataattga aaacgcgtag
     5221 ttataatcgc tgaggtaata tttaaaatca ttttcaaatg attcacagtt aatttgcgac
     5281 aatataattt tattttcaca taaactagac gccttgtcgt cttcttcttc gtattccttc
     5341 tctttttcat ttttctcttc ataaaaatta acatagttat tatcgtatcc atatatgtat
     5401 ctatcgtata gagtaaattt tttgttgtca taaatatata tgtctttttt aatggggtgt
     5461 atagtaccgc tgcgcatagt ttttctgtaa tttacaacag tgctattttc tggtagttct
     5521 tcggagtgtg ttgctttaat tattaaattt atataatcaa tgaatttggg atcgtcggtt
     5581 ttatacaata tgttgccggc atagtacgca gcttcttcta gttcaattac accatttttt
     5641 agcagcaccg gattaacata actttccaaa atgttgtacg aaccgttaaa caaaaacagt
     5701 tcacctccct tttctatact attgtctgcg agcagttgtt tgttgttaaa aataacagcc
     5761 attgtaatga gacgcacaaa ctaatatcac aaactggaaa tgtctatcaa tatatagttg
     5821 ctgatggccg gcctattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5881 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     5941 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     6001 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     6061 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     6121 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     6181 cgctattacc atgctgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     6241 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     6301 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     6361 tagg
//
