LOCUS       sequence-375290-        6559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces an mTurquoise-antiFRET-mScarlet3 fusion protein in bacteria
            and in mamallian cells it produces 1:1 unfused mTurquoise2 and 
            mScarlet3 FPs for ratiometric analysis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6559)
  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 6559)
  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: pDRESS_mTurquoise2_spatial-linker-P2A_mScarlet3
FEATURES             Location/Qualifiers
     source          1..6559
                     /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             join(424..426,427..429,430..1140)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(469..490)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(730..749)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1077..1098
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1261..1317
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     regulatory      1336..1345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1342..2028
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     1740..1759
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2130..2149)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2195..2250
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2249..2268)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2338..2385
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2398..3103
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2486..2858
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2515..3103)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3112..3145)
                     /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(3313..3900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3392..3411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3913..4728)
                     /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     4636..4655
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_recomb     4971..5798
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4971..5095
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5133..5738)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5866..6169
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6171..6373
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6324..6344
                     /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 gccggtcgcc
      421 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      481 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      541 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      601 accctcgtga ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg
      661 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      721 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      781 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      841 cacaagctgg agtacaacta ctttagcgac aacgtctata tcaccgccga caagcagaag
      901 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
      961 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1021 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1081 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctctacaag
     1141 tcgaccagat ctctggccga ggcggcagct aaggaggccg cggcgaagga ggcagccgcg
     1201 aaagaagcag ccgcaaagga agcggccgcc aaggctgctg caagatctct cggatcgggt
     1261 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggaccttcg
     1321 ctagcgctac cggtcgccac catggatagc accgaggcag tgatcaagga gttcatgcgg
     1381 ttcaaggtgc acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     1441 gagggccgcc cctacgaggg cacccagacc gccaagctga gggtgaccaa gggtggcccc
     1501 ctgcccttct cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacg
     1561 aagcaccccg ccgacatccc cgactactgg aagcagtcct tccccgaggg cttcaagtgg
     1621 gagcgcgtga tgaacttcga ggacggcggc gccgtgtccg tggcccagga cacctccctg
     1681 gaggacggca ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc
     1741 cccgtaatgc agaagaagac aatgggctgg gaagcatcca ccgagcggtt gtaccccgag
     1801 gacgtcgtgc tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac
     1861 ctggcggact tcaagaccac ctacagggcc aagaagcccg tgcagatgcc cggcgccttc
     1921 aacatcgacc gcaagttgga catcacatcc cacaacgagg actacaccgt ggtggaacag
     1981 tacgaacgct ccgtggcccg ccactccacc ggcggctccg gtggctccta actgtacaag
     2041 taagtgatta acctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     2101 ccctggctca caaataccac tgagatcgat ctttttccct ctgccaaaaa ttatggggac
     2161 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     2221 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     2281 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgtaacta
     2341 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa agcatgcgga
     2401 ggaaattctc cttgaagttt ccctggtgtt caaagtaaag gagtttgcac cagacgcacc
     2461 tctgttcact ggtccggcgt attaaaacac gatacattgt tattagtaca tttattaagc
     2521 gctagattct gtgcgttgtt gatttacaga caattgttgt acgtatttta ataattcatt
     2581 aaatttataa tctttagggt ggtatgttag agcgaaaatc aaatgatttt cagcgtcttt
     2641 atatctgaat ttaaatatta aatcctcaat agatttgtaa aataggtttc gattagtttc
     2701 aaacaagggt tgtttttccg aaccgatggc tggactatct aatggatttt cgctcaacgc
     2761 cacaaaactt gccaaatctt gtagcagcaa tctagctttg tcgatattcg tttgtgtttt
     2821 gttttgtaat aaaggttcga cgtcgttcaa aatattatgc gcttttgtat ttctttcatc
     2881 actgtcgtta gtatacaatt gactcgacgt aaacacgtta aatagagctt ggacatattt
     2941 aacatcgggc gtgttagctt tattaggccg attatcgtcg tcgtcccaac cctcgtcgtt
     3001 agaagttgct tccgaagacg attttgccat agccacacga cgcctattaa ttgtgtcggc
     3061 taacacgtcc gcgatcaaat ttgtagttga gctttttgga attgcgatcg cataacttcg
     3121 tatagcatac attatacgaa gttataagct cggaacgctg cgctcggtcg ttcggctgcg
     3181 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3241 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3301 gttgctggcg tttttccata ggctccgccc cctgacgagc atcacaaaaa tcgacgctca
     3361 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3421 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3481 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3541 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3601 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3661 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3721 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3781 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3841 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3901 gaagatcctt tgttagaaaa actcatcgag catcaagtga aactgcaatt tattcatatc
     3961 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     4021 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     4081 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     4141 atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg
     4201 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcaccaacca aaccgttatt
     4261 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ccgttaaaag gacaattaca
     4321 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     4381 tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag
     4441 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     4501 cgtcagccag tttagcctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     4561 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc
     4621 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     4681 atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcatac tcttcctttt
     4741 tcaatattat tgaagcattt atcagggtta ttgtctcatg tccgcgcgtt tcctgcatct
     4801 tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tataggaaga ggtttatact
     4861 aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat gtttaatcaa
     4921 ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca tgcatctttt
     4981 aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa ctgtcgacaa
     5041 gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt attgaataat
     5101 aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa cgcaacaaga
     5161 acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag gtaatattta
     5221 aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt ttcacataaa
     5281 ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt ctcttcataa
     5341 aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt aaattttttg
     5401 ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg catagttttt
     5461 ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc tttaattatt
     5521 aaatttatat aatcaatgaa tttgggatcg tcggttttat acaatatgtt gccggcatag
     5581 tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt aacataactt
     5641 tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc tatactattg
     5701 tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg cacaaactaa
     5761 tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggcct attaatagta
     5821 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5881 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5941 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6001 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     6061 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     6121 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6181 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6241 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6301 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6361 tataagcaga cgtcgtttag tgaaccgtca gatcactaga tgctttattg cggtagttta
     6421 tcacagttaa attgctaacg ccagtctcga acttaacgtg cagaagttgg tcgtgaggca
     6481 ctgggcaggt aagtatcggg ccctttgtgc ggggggagcg gctcggggct gtccgcgggg
     6541 ggacggctgc cttcggggg
//
