LOCUS       sequence-375295-        5641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression vector for bacteria and mammalian cells producing 
            mScarlet3 red fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5641)
  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 5641)
  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_mScarlet3
FEATURES             Location/Qualifiers
     source          1..5641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     238..260
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     322..341
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        383..501
                     /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             576..593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             603..620
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             645..1331
                     /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     1043..1062
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1433..1452)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1498..1553
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1552..1571)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      1641..1688
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     1701..2406
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           1789..2161
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(1818..2406)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2415..2448)
                     /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(2616..3203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2695..2714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3216..4031)
                     /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     3939..3958
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_recomb     4274..5101
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4274..4398
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4436..5041)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5169..5472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5474..5641,1..35)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(5627..5641,1..6)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggcgtgtacg gtgggaggtc tatataagca gacgtcgttt agtgaaccgt cagatcacta
       61 gatgctttat tgcggtagtt tatcacagtt aaattgctaa cgccagtctc gaacttaacg
      121 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatcg ggccctttgt gcggggggag
      181 cggctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
      241 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
      301 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
      361 aagaattgga tcggaccgaa atcaccacaa ttcagcaaat tgtgaacatc atcacgttca
      421 tctttccctg gttgccaaag gcccattttc ctgtcagtaa cgagaaggtc gcgaattcag
      481 gcgcttttta gactggtcgt agggagacca caacggtttc cctctagaaa taattttgtt
      541 taactataag aaggagatat accatgcggg gttctcatca tcaccaccat cacagcagcg
      601 gcctggtgcc gcgcggcagc tcgctagcgc taccggtcgc caccatggat agcaccgagg
      661 cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg
      721 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
      781 tgagggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca
      841 tgtacggctc cagggccttc acgaagcacc ccgccgacat ccccgactac tggaagcagt
      901 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtgt
      961 ccgtggccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg
     1021 gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcat
     1081 ccaccgagcg gttgtacccc gaggacgtcg tgctgaaggg cgacattaag atggccctgc
     1141 gcctgaagga cggcggccgc tacctggcgg acttcaagac cacctacagg gccaagaagc
     1201 ccgtgcagat gcccggcgcc ttcaacatcg accgcaagtt ggacatcaca tcccacaacg
     1261 aggactacac cgtggtggaa cagtacgaac gctccgtggc ccgccactcc accggcggct
     1321 ccggtggctc ctaactgtac aagtaagtga ttaacctcag gtgcaggctg cctatcagaa
     1381 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc gatctttttc
     1441 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     1501 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     1561 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     1621 caacatatgc ccatatgtaa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1681 gttttttgct gaaagcatgc ggaggaaatt ctccttgaag tttccctggt gttcaaagta
     1741 aaggagtttg caccagacgc acctctgttc actggtccgg cgtattaaaa cacgatacat
     1801 tgttattagt acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt
     1861 tgtacgtatt ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa
     1921 atcaaatgat tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg
     1981 taaaataggt ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta
     2041 tctaatggat tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct
     2101 ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta
     2161 tgcgcttttg tatttctttc atcactgtcg ttagtataca attgactcga cgtaaacacg
     2221 ttaaatagag cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg
     2281 tcgtcgtccc aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca
     2341 cgacgcctat taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt
     2401 ggaattgcga tcgcataact tcgtatagca tacattatac gaagttataa gctcggaacg
     2461 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg ccccctgacg
     2641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3121 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3181 cgcgcagaaa aaaaggatct caagaagatc ctttgttaga aaaactcatc gagcatcaag
     3241 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     3301 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     3361 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     3421 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     3481 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     3541 ctcgcaccaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     3601 tcgccgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     3661 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     3721 ttccctggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     3781 atggtcggaa gaggcataaa ttccgtcagc cagtttagcc tgaccatctc atctgtaaca
     3841 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     3901 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3961 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     4021 atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4081 atgtccgcgc gtttcctgca tcttttaatc aaatcccaag atgtgtataa acgcgccggt
     4141 atgtatagga agaggtttat actaaactgt tacattgcaa acgtggtttc gtgtgccaag
     4201 tgtgaaaacc gatgtttaat caaggctctg acgcatttct acaaccacga ctccaagtgt
     4261 gtgggtgaag tcatgcatct tttaatcaaa tcccaagatg tgtataaacc accaaactgc
     4321 caaaaaatga aaactgtcga caagctctgt ccgtttgctg gcaactgcaa gggtctcaat
     4381 cctatttgta attattgaat aataaaacaa ttataaatgt caaatttgtt ttttattaac
     4441 gatacaaacc aaacgcaaca agaacatttg tagtattatc tataattgaa aacgcgtagt
     4501 tataatcgct gaggtaatat ttaaaatcat tttcaaatga ttcacagtta atttgcgaca
     4561 atataatttt attttcacat aaactagacg ccttgtcgtc ttcttcttcg tattccttct
     4621 ctttttcatt tttctcttca taaaaattaa catagttatt atcgtatcca tatatgtatc
     4681 tatcgtatag agtaaatttt ttgttgtcat aaatatatat gtctttttta atggggtgta
     4741 tagtaccgct gcgcatagtt tttctgtaat ttacaacagt gctattttct ggtagttctt
     4801 cggagtgtgt tgctttaatt attaaattta tataatcaat gaatttggga tcgtcggttt
     4861 tatacaatat gttgccggca tagtacgcag cttcttctag ttcaattaca ccatttttta
     4921 gcagcaccgg attaacataa ctttccaaaa tgttgtacga accgttaaac aaaaacagtt
     4981 cacctccctt ttctatacta ttgtctgcga gcagttgttt gttgttaaaa ataacagcca
     5041 ttgtaatgag acgcacaaac taatatcaca aactggaaat gtctatcaat atatagttgc
     5101 tgatggccgg cctattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5161 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5221 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5281 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5341 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5401 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
     5461 gctattacca tgctgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
     5521 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     5581 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
     5641 a
//
