LOCUS       sequence-262638-        6556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6556)
  AUTHORS   Bindels DS, Postma M, Haarbosch L, van Weeren L, Gadella TWJ Jr
  TITLE     Multiparameter screening method for developing optimized 
            red-fluorescent proteins.
  JOURNAL   Nat Protoc. 2020 Feb;15(2):450-478. doi: 10.1038/s41596-019-0250-7.
  PUBMED    31942080
REFERENCE   2  (bases 1 to 6556)
  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"; journalName: 
            "Nat Protoc"; date: "2020-02"; volume: "15"; issue: "2"; pages: 
            "450-478"
COMMENT     Sequence Label: pDRESS_mTurquoise2_spatial-linker-P2A_mScarlet
FEATURES             Location/Qualifiers
     source          1..6556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     181..1008
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             181..305
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(343..948)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1076..1379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1381..1583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1534..1554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1786..1808
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1870..1889
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        1931..2049
                     /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             2124..2141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2151..2168
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      2187..2196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2193..2195,2196..2198,2199..2909)
                     /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(2238..2259)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2499..2518)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2846..2867
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3030..3086
                     /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      3105..3114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3111..3806
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3244..3262)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3512..3531
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3896..3915)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3961..4016
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4015..4034)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      4104..4151
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     4164..4869
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           4252..4624
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(4281..4869)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(4878..4911)
                     /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(5079..5666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5158..5177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5679..6494)
                     /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     6402..6421
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tataggaaga
       61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
      121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
      181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
      241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
      301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
      361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
      421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
      481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
      541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
      601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
      661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
      721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttat acaatatgtt
      781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
      841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
      901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
      961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggcct
     1021 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     1081 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1141 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1201 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc
     1261 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1321 ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     1381 tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc
     1441 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1501 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
     1561 gggaggtcta tataagcaga cgtcgtttag tgaaccgtca gatcactaga tgctttattg
     1621 cggtagttta tcacagttaa attgctaacg ccagtctcga acttaacgtg cagaagttgg
     1681 tcgtgaggca ctgggcaggt aagtatcggg ccctttgtgc ggggggagcg gctcggggct
     1741 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1801 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1861 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc
     1921 ggaccgaaat caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt
     1981 tgccaaaggc ccattttcct gtcagtaacg agaaggtcgc gaattcaggc gctttttaga
     2041 ctggtcgtag ggagaccaca acggtttccc tctagaaata attttgttta actataagaa
     2101 ggagatatac catgcggggt tctcatcatc accaccatca cagcagcggc ctggtgccgc
     2161 gcggcagctc gctagcgctg ccggtcgcca ccatggtgag caagggcgag gagctgttca
     2221 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2281 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2341 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgtcc tggggcgtgc
     2401 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2461 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2521 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2581 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac tttagcgaca
     2641 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     2701 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2761 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     2821 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2881 tcactctcgg catggacgag ctctacaagt cgaccagatc tctggccgag gcggcagcta
     2941 aggaggccgc ggcgaaggag gcagccgcga aagaagcagc cgcaaaggaa gcggccgcca
     3001 aggctgctgc aagatctctc ggatcgggtg ctactaactt cagcctgctg aagcaggctg
     3061 gagacgtgga ggagaaccct ggaccttcgc tagcgctacc ggtcgccacc atggtgagca
     3121 agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga
     3181 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     3241 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc
     3301 ctcagttcat gtacggctcc agggccttca ccaagcaccc cgccgacatc cccgactact
     3361 ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     3421 gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtga
     3481 agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct
     3541 gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga
     3601 tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc acctacaagg
     3661 ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg gacatcacct
     3721 cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca
     3781 ccggcggcat ggacgagctg tacaagtaag tgattaacct caggtgcagg ctgcctatca
     3841 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atcgatcttt
     3901 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     3961 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     4021 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     4081 tggcaacata tgcccatatg taactagcat aaccccttgg ggcctctaaa cgggtcttga
     4141 ggggtttttt gctgaaagca tgcggaggaa attctccttg aagtttccct ggtgttcaaa
     4201 gtaaaggagt ttgcaccaga cgcacctctg ttcactggtc cggcgtatta aaacacgata
     4261 cattgttatt agtacattta ttaagcgcta gattctgtgc gttgttgatt tacagacaat
     4321 tgttgtacgt attttaataa ttcattaaat ttataatctt tagggtggta tgttagagcg
     4381 aaaatcaaat gattttcagc gtctttatat ctgaatttaa atattaaatc ctcaatagat
     4441 ttgtaaaata ggtttcgatt agtttcaaac aagggttgtt tttccgaacc gatggctgga
     4501 ctatctaatg gattttcgct caacgccaca aaacttgcca aatcttgtag cagcaatcta
     4561 gctttgtcga tattcgtttg tgttttgttt tgtaataaag gttcgacgtc gttcaaaata
     4621 ttatgcgctt ttgtatttct ttcatcactg tcgttagtat acaattgact cgacgtaaac
     4681 acgttaaata gagcttggac atatttaaca tcgggcgtgt tagctttatt aggccgatta
     4741 tcgtcgtcgt cccaaccctc gtcgttagaa gttgcttccg aagacgattt tgccatagcc
     4801 acacgacgcc tattaattgt gtcggctaac acgtccgcga tcaaatttgt agttgagctt
     4861 tttggaattg cgatcgcata acttcgtata gcatacatta tacgaagtta taagctcgga
     4921 acgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4981 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgccccctg
     5101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgtt agaaaaactc atcgagcatc
     5701 aagtgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     5761 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     5821 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     5881 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     5941 agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     6001 tcactcgcac caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     6061 cgatcgccgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     6121 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     6181 gttttccctg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     6241 ttgatggtcg gaagaggcat aaattccgtc agccagttta gcctgaccat ctcatctgta
     6301 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     6361 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     6421 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt
     6481 tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6541 ctcatgtccg cgcgtt
//
