LOCUS       sequence-375312-        6355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression vector for bacteria and mammalian cells producing 
            FusionRed-SYFP2 fusion protein for FRET analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6355)
  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 6355)
  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_FusionRed-SYFP2
FEATURES             Location/Qualifiers
     source          1..6355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..39
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     101..120
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        162..280
                     /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             355..372
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             382..399
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      418..427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             424..1119
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDLYK"
     CDS             join(1120..1122,1123..1125,1126..1836)
                     /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(1165..1186)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1426..1445)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1773..1794
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1926..1945)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1991..2046
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2045..2064)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2134..2181
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2194..2899
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2282..2654
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2311..2899)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2908..2941)
                     /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(3109..3696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3188..3207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3709..4524)
                     /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     4432..4451
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_recomb     4767..5594
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4767..4891
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4929..5534)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5662..5965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5967..6169
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6120..6140
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
       61 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
      121 ctgtctcatc attttggcaa agaattggat cggaccgaaa tcaccacaat tcagcaaatt
      181 gtgaacatca tcacgttcat ctttccctgg ttgccaaagg cccattttcc tgtcagtaac
      241 gagaaggtcg cgaattcagg cgctttttag actggtcgta gggagaccac aacggtttcc
      301 ctctagaaat aattttgttt aactataaga aggagatata ccatgcgggg ttctcatcat
      361 caccaccatc acagcagcgg cctggtgccg cgcggcagct cgctagcgct accggtcgcc
      421 accatggtga gcgagctgat taaggagaac atgcccatga agctctacat ggagggcacc
      481 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
      541 cagaccatga gaatcaaggt cgtcgagggc ggccctctcc ccttcgcctt cgacatcctg
      601 gctaccagct tcatgtacgg cagcagaacc ttcatcaagc accctccggg catccccgac
      661 ttctttaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
      721 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
      781 gtcaaggtta gaggggtgaa cttcccagcc aacggccctg tgatgcagaa gaaaacactc
      841 ggctgggagg cctccaccga gacgatgtac cccgctgacg gcggcctgga aggcgcatgt
      901 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaaccttga gaccacatac
      961 agatccaaga aacccgctac gaacctcaag atgcccggcg tctacaacgt ggaccacaga
     1021 ctggaaagaa tcaaggaggc cgacgatgag acctacgtcg agcagcacga ggtggctgtg
     1081 gccagatact ctactggtgg cgctggtgat ctgtacaaga tggtgagcaa gggcgaggag
     1141 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1201 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1261 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1321 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1381 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1441 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1501 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1561 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1621 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1681 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1741 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1801 gccgggatca ctctcggcat ggacgagctc tacaagtaag tgattaacct caggtgcagg
     1861 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     1921 atcgatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     1981 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     2041 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     2101 gtttagagtt tggcaacata tgcccatatg taactagcat aaccccttgg ggcctctaaa
     2161 cgggtcttga ggggtttttt gctgaaagca tgcggaggaa attctccttg aagtttccct
     2221 ggtgttcaaa gtaaaggagt ttgcaccaga cgcacctctg ttcactggtc cggcgtatta
     2281 aaacacgata cattgttatt agtacattta ttaagcgcta gattctgtgc gttgttgatt
     2341 tacagacaat tgttgtacgt attttaataa ttcattaaat ttataatctt tagggtggta
     2401 tgttagagcg aaaatcaaat gattttcagc gtctttatat ctgaatttaa atattaaatc
     2461 ctcaatagat ttgtaaaata ggtttcgatt agtttcaaac aagggttgtt tttccgaacc
     2521 gatggctgga ctatctaatg gattttcgct caacgccaca aaacttgcca aatcttgtag
     2581 cagcaatcta gctttgtcga tattcgtttg tgttttgttt tgtaataaag gttcgacgtc
     2641 gttcaaaata ttatgcgctt ttgtatttct ttcatcactg tcgttagtat acaattgact
     2701 cgacgtaaac acgttaaata gagcttggac atatttaaca tcgggcgtgt tagctttatt
     2761 aggccgatta tcgtcgtcgt cccaaccctc gtcgttagaa gttgcttccg aagacgattt
     2821 tgccatagcc acacgacgcc tattaattgt gtcggctaac acgtccgcga tcaaatttgt
     2881 agttgagctt tttggaattg cgatcgcata acttcgtata gcatacatta tacgaagtta
     2941 taagctcgga acgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3121 ccgccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3181 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3241 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3301 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3361 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3421 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3481 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3541 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3601 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3661 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgtt agaaaaactc
     3721 atcgagcatc aagtgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     3781 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     3841 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     3901 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     3961 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     4021 gtcatcaaaa tcactcgcac caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     4081 acgaaatacg cgatcgccgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     4141 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     4201 ctggaatgct gttttccctg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     4261 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gcctgaccat
     4321 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     4381 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     4441 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga
     4501 cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca
     4561 gggttattgt ctcatgtccg cgcgtttcct gcatctttta atcaaatccc aagatgtgta
     4621 taaacgcgcc ggtatgtata ggaagaggtt tatactaaac tgttacattg caaacgtggt
     4681 ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct ctgacgcatt tctacaacca
     4741 cgactccaag tgtgtgggtg aagtcatgca tcttttaatc aaatcccaag atgtgtataa
     4801 accaccaaac tgccaaaaaa tgaaaactgt cgacaagctc tgtccgtttg ctggcaactg
     4861 caagggtctc aatcctattt gtaattattg aataataaaa caattataaa tgtcaaattt
     4921 gttttttatt aacgatacaa accaaacgca acaagaacat ttgtagtatt atctataatt
     4981 gaaaacgcgt agttataatc gctgaggtaa tatttaaaat cattttcaaa tgattcacag
     5041 ttaatttgcg acaatataat tttattttca cataaactag acgccttgtc gtcttcttct
     5101 tcgtattcct tctctttttc atttttctct tcataaaaat taacatagtt attatcgtat
     5161 ccatatatgt atctatcgta tagagtaaat tttttgttgt cataaatata tatgtctttt
     5221 ttaatggggt gtatagtacc gctgcgcata gtttttctgt aatttacaac agtgctattt
     5281 tctggtagtt cttcggagtg tgttgcttta attattaaat ttatataatc aatgaatttg
     5341 ggatcgtcgg ttttatacaa tatgttgccg gcatagtacg cagcttcttc tagttcaatt
     5401 acaccatttt ttagcagcac cggattaaca taactttcca aaatgttgta cgaaccgtta
     5461 aacaaaaaca gttcacctcc cttttctata ctattgtctg cgagcagttg tttgttgtta
     5521 aaaataacag ccattgtaat gagacgcaca aactaatatc acaaactgga aatgtctatc
     5581 aatatatagt tgctgatggc cggcctatta atagtaatca attacggggt cattagttca
     5641 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5701 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5761 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5821 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     5881 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     5941 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacacca atgggcgtgg
     6001 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6061 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     6121 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagacgtc gtttagtgaa
     6181 ccgtcagatc actagatgct ttattgcggt agtttatcac agttaaattg ctaacgccag
     6241 tctcgaactt aacgtgcaga agttggtcgt gaggcactgg gcaggtaagt atcgggccct
     6301 ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac ggctgccttc ggggg
//
