LOCUS       sequence-375301-        5641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression vector for bacteria and mammalian cells producing 
            mScarlet-I3 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_mScarlet-I3
FEATURES             Location/Qualifiers
     source          1..5641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     242..264
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     326..345
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        387..505
                     /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             580..597
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             607..624
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      643..652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             649..1335
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGGT
                     VSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     complement(779..797)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1437..1456)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1502..1557
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1556..1575)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      1645..1692
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     1705..2410
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           1793..2165
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(1822..2410)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2419..2452)
                     /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(2620..3207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2699..2718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3220..4035)
                     /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     3943..3962
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_recomb     4278..5105
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4278..4402
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4440..5045)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5173..5476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5478..5641,1..39)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(5631..5641,1..10)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggtaggcgtg tacggtggga ggtctatata agcagacgtc gtttagtgaa ccgtcagatc
       61 actagatgct ttattgcggt agtttatcac agttaaattg ctaacgccag tctcgaactt
      121 aacgtgcaga agttggtcgt gaggcactgg gcaggtaagt atcgggccct ttgtgcgggg
      181 ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg
      241 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc
      301 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
      361 ggcaaagaat tggatcggac cgaaatcacc acaattcagc aaattgtgaa catcatcacg
      421 ttcatctttc cctggttgcc aaaggcccat tttcctgtca gtaacgagaa ggtcgcgaat
      481 tcaggcgctt tttagactgg tcgtagggag accacaacgg tttccctcta gaaataattt
      541 tgtttaacta taagaaggag atataccatg cggggttctc atcatcacca ccatcacagc
      601 agcggcctgg tgccgcgcgg cagctcgcta gcgctaccgg tcgccaccat ggatagcacc
      661 gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc
      721 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
      781 aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag
      841 ttcatgtacg gctccagggc cttcatcaag caccccgccg acatccccga ctactggaag
      901 cagtccttcc ccgagggctt caagtgggag cgcgtgatga tcttcgagga cggcggcacc
      961 gtgtccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc
     1021 cgcggcggca acttccctcc tgacggcccc gtaatgcaga agaggacaat gggctgggaa
     1081 gcatccaccg agcggttgta ccccgaggac gtcgtgctga agggcgacat taagatggcc
     1141 ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag
     1201 aagcccgtgc agatgcccgg cgccttcaac atcgaccgca agttggacat cacctcccac
     1261 aacgaggact acaccgtggt ggaacagtac gaacgctccg tggcccgcca ctccaccggc
     1321 ggctccggtg gctcctaact gtacaagtaa gtgattaacc tcaggtgcag gctgcctatc
     1381 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatcgatctt
     1441 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     1501 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     1561 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     1621 ttggcaacat atgcccatat gtaactagca taaccccttg gggcctctaa acgggtcttg
     1681 aggggttttt tgctgaaagc atgcggagga aattctcctt gaagtttccc tggtgttcaa
     1741 agtaaaggag tttgcaccag acgcacctct gttcactggt ccggcgtatt aaaacacgat
     1801 acattgttat tagtacattt attaagcgct agattctgtg cgttgttgat ttacagacaa
     1861 ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt atgttagagc
     1921 gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat cctcaataga
     1981 tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac cgatggctgg
     2041 actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta gcagcaatct
     2101 agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat
     2161 attatgcgct tttgtatttc tttcatcact gtcgttagta tacaattgac tcgacgtaaa
     2221 cacgttaaat agagcttgga catatttaac atcgggcgtg ttagctttat taggccgatt
     2281 atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt ttgccatagc
     2341 cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg tagttgagct
     2401 ttttggaatt gcgatcgcat aacttcgtat agcatacatt atacgaagtt ataagctcgg
     2461 aacgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccccct
     2641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3181 attacgcgca gaaaaaaagg atctcaagaa gatcctttgt tagaaaaact catcgagcat
     3241 caagtgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     3301 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     3361 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     3421 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     3481 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     3541 atcactcgca ccaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     3601 gcgatcgccg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     3661 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     3721 tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     3781 cttgatggtc ggaagaggca taaattccgt cagccagttt agcctgacca tctcatctgt
     3841 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     3901 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     3961 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg
     4021 ttgaatatgg ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4081 tctcatgtcc gcgcgtttcc tgcatctttt aatcaaatcc caagatgtgt ataaacgcgc
     4141 cggtatgtat aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc
     4201 caagtgtgaa aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa
     4261 gtgtgtgggt gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa
     4321 ctgccaaaaa atgaaaactg tcgacaagct ctgtccgttt gctggcaact gcaagggtct
     4381 caatcctatt tgtaattatt gaataataaa acaattataa atgtcaaatt tgttttttat
     4441 taacgataca aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg
     4501 tagttataat cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc
     4561 gacaatataa ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc
     4621 ttctcttttt catttttctc ttcataaaaa ttaacatagt tattatcgta tccatatatg
     4681 tatctatcgt atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg
     4741 tgtatagtac cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt
     4801 tcttcggagt gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg
     4861 gttttataca atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt
     4921 tttagcagca ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac
     4981 agttcacctc ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca
     5041 gccattgtaa tgagacgcac aaactaatat cacaaactgg aaatgtctat caatatatag
     5101 ttgctgatgg ccggcctatt aatagtaatc aattacgggg tcattagttc atagcccata
     5161 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5221 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5281 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     5341 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     5401 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
     5461 catcgctatt accatgctga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt
     5521 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     5581 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
     5641 c
//
