LOCUS       sequence-375307-        6367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression vector for bacteria and mammalian cells producing 
            mCherry-SYFP2 fusion protein for FRET analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6367)
  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 6367)
  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_mCherry-SYFP2
FEATURES             Location/Qualifiers
     source          1..6367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..41
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     103..122
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        164..282
                     /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             357..374
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             384..401
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      420..429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             426..1133
                     /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(571..589)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     839..858
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(1134..1136,1137..1139,1140..1850)
                     /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(1179..1200)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1440..1459)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1787..1808
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1940..1959)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2005..2060
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2059..2078)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2148..2195
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2208..2913
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2296..2668
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2325..2913)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2922..2955)
                     /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(3123..3710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3202..3221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3723..4538)
                     /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     4446..4465
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_recomb     4781..5608
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4781..4905
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4943..5548)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5676..5979
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5981..6183
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6134..6154
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
       61 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
      121 tgctgtctca tcattttggc aaagaattgg atcggaccga aatcaccaca attcagcaaa
      181 ttgtgaacat catcacgttc atctttccct ggttgccaaa ggcccatttt cctgtcagta
      241 acgagaaggt cgcgaattca ggcgcttttt agactggtcg tagggagacc acaacggttt
      301 ccctctagaa ataattttgt ttaactataa gaaggagata taccatgcgg ggttctcatc
      361 atcaccacca tcacagcagc ggcctggtgc cgcgcggcag ctcgctagcg ctaccggtcg
      421 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      481 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      541 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
      601 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      661 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      721 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      781 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
      841 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
      901 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
      961 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1021 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1081 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagatggtga
     1141 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1201 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1261 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1321 ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1381 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1441 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1501 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1561 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     1621 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     1681 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1741 gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1801 agttcgtgac cgccgccggg atcactctcg gcatggacga gctctacaag taagtgatta
     1861 acctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     1921 caaataccac tgagatcgat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     1981 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     2041 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     2101 agaatgagta tttggtttag agtttggcaa catatgccca tatgtaacta gcataacccc
     2161 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa agcatgcgga ggaaattctc
     2221 cttgaagttt ccctggtgtt caaagtaaag gagtttgcac cagacgcacc tctgttcact
     2281 ggtccggcgt attaaaacac gatacattgt tattagtaca tttattaagc gctagattct
     2341 gtgcgttgtt gatttacaga caattgttgt acgtatttta ataattcatt aaatttataa
     2401 tctttagggt ggtatgttag agcgaaaatc aaatgatttt cagcgtcttt atatctgaat
     2461 ttaaatatta aatcctcaat agatttgtaa aataggtttc gattagtttc aaacaagggt
     2521 tgtttttccg aaccgatggc tggactatct aatggatttt cgctcaacgc cacaaaactt
     2581 gccaaatctt gtagcagcaa tctagctttg tcgatattcg tttgtgtttt gttttgtaat
     2641 aaaggttcga cgtcgttcaa aatattatgc gcttttgtat ttctttcatc actgtcgtta
     2701 gtatacaatt gactcgacgt aaacacgtta aatagagctt ggacatattt aacatcgggc
     2761 gtgttagctt tattaggccg attatcgtcg tcgtcccaac cctcgtcgtt agaagttgct
     2821 tccgaagacg attttgccat agccacacga cgcctattaa ttgtgtcggc taacacgtcc
     2881 gcgatcaaat ttgtagttga gctttttgga attgcgatcg cataacttcg tatagcatac
     2941 attatacgaa gttataagct cggaacgctg cgctcggtcg ttcggctgcg gcgagcggta
     3001 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3061 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3121 tttttccata ggctccgccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3181 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3241 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3301 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3361 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3421 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3481 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3541 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3601 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3661 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3721 tgttagaaaa actcatcgag catcaagtga aactgcaatt tattcatatc aggattatca
     3781 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     3841 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     3901 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     3961 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     4021 cagccattac gctcgtcatc aaaatcactc gcaccaacca aaccgttatt cattcgtgat
     4081 tgcgcctgag cgagacgaaa tacgcgatcg ccgttaaaag gacaattaca aacaggaatc
     4141 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     4201 tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag taaccatgca
     4261 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     4321 tttagcctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     4381 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     4441 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     4501 ggcctcgagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     4561 tgaagcattt atcagggtta ttgtctcatg tccgcgcgtt tcctgcatct tttaatcaaa
     4621 tcccaagatg tgtataaacg cgccggtatg tataggaaga ggtttatact aaactgttac
     4681 attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat gtttaatcaa ggctctgacg
     4741 catttctaca accacgactc caagtgtgtg ggtgaagtca tgcatctttt aatcaaatcc
     4801 caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa ctgtcgacaa gctctgtccg
     4861 tttgctggca actgcaaggg tctcaatcct atttgtaatt attgaataat aaaacaatta
     4921 taaatgtcaa atttgttttt tattaacgat acaaaccaaa cgcaacaaga acatttgtag
     4981 tattatctat aattgaaaac gcgtagttat aatcgctgag gtaatattta aaatcatttt
     5041 caaatgattc acagttaatt tgcgacaata taattttatt ttcacataaa ctagacgcct
     5101 tgtcgtcttc ttcttcgtat tccttctctt tttcattttt ctcttcataa aaattaacat
     5161 agttattatc gtatccatat atgtatctat cgtatagagt aaattttttg ttgtcataaa
     5221 tatatatgtc ttttttaatg gggtgtatag taccgctgcg catagttttt ctgtaattta
     5281 caacagtgct attttctggt agttcttcgg agtgtgttgc tttaattatt aaatttatat
     5341 aatcaatgaa tttgggatcg tcggttttat acaatatgtt gccggcatag tacgcagctt
     5401 cttctagttc aattacacca ttttttagca gcaccggatt aacataactt tccaaaatgt
     5461 tgtacgaacc gttaaacaaa aacagttcac ctcccttttc tatactattg tctgcgagca
     5521 gttgtttgtt gttaaaaata acagccattg taatgagacg cacaaactaa tatcacaaac
     5581 tggaaatgtc tatcaatata tagttgctga tggccggcct attaatagta atcaattacg
     5641 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5701 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5761 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5821 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
     5881 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
     5941 tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac
     6001 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     6061 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac
     6121 cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     6181 cgtcgtttag tgaaccgtca gatcactaga tgctttattg cggtagttta tcacagttaa
     6241 attgctaacg ccagtctcga acttaacgtg cagaagttgg tcgtgaggca ctgggcaggt
     6301 aagtatcggg ccctttgtgc ggggggagcg gctcggggct gtccgcgggg ggacggctgc
     6361 cttcggg
//
