LOCUS       sequence-492366-        5638 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  Dual expression vector for bacteria and mammalian cells producing 
            mScarlet-3-H red fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5638)
  TITLE     mScarlet3-H plasmids
REFERENCE   2  (bases 1 to 5638)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pDx_mScarlet3-H
FEATURES             Location/Qualifiers
     source          1..5638
                     /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             2193..2879
                     /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
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     2591..2610
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2978..2997)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3043..3098
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3097..3116)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3186..3233
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3246..3951
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3334..3706
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3363..3951)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3960..3993)
                     /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(4161..4748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4240..4259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4761..5576)
                     /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     5484..5503
                     /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 gctagcgcta ccggtcgcca ccatggatag caccgaggca gtgatcaagg
     2221 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
     2281 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg agggtgacca
     2341 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2401 gggccttcac gaagcacccc gccgacatcc ccgactactg gaagcagtcc ttccccgagg
     2461 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgtcc gtggcccagg
     2521 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     2581 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcatcc accgagcggt
     2641 tgtaccccga ggacgtcgtg ctgaagggcg acattaagca cgccctgcgc ctgaaggacg
     2701 gcggccgcta cctggcggac ttcaagacca cctacagggc caagaagccc gtgcagatgc
     2761 ccggcgcctt caacatcgac cgcaagttgg acatcacatc ccacaacgag gactacaccg
     2821 tggtggaaca gtacgaacgc tccgtggccc gccactccac cggcggctcc ggtggctcct
     2881 tgtacaagta agtgattaac ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     2941 ggccaatgcc ctggctcaca aataccactg agatcgatct ttttccctct gccaaaaatt
     3001 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     3061 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     3121 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     3181 tgtaactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     3241 catgcggagg aaattctcct tgaagtttcc ctggtgttca aagtaaagga gtttgcacca
     3301 gacgcacctc tgttcactgg tccggcgtat taaaacacga tacattgtta ttagtacatt
     3361 tattaagcgc tagattctgt gcgttgttga tttacagaca attgttgtac gtattttaat
     3421 aattcattaa atttataatc tttagggtgg tatgttagag cgaaaatcaa atgattttca
     3481 gcgtctttat atctgaattt aaatattaaa tcctcaatag atttgtaaaa taggtttcga
     3541 ttagtttcaa acaagggttg tttttccgaa ccgatggctg gactatctaa tggattttcg
     3601 ctcaacgcca caaaacttgc caaatcttgt agcagcaatc tagctttgtc gatattcgtt
     3661 tgtgttttgt tttgtaataa aggttcgacg tcgttcaaaa tattatgcgc ttttgtattt
     3721 ctttcatcac tgtcgttagt atacaattga ctcgacgtaa acacgttaaa tagagcttgg
     3781 acatatttaa catcgggcgt gttagcttta ttaggccgat tatcgtcgtc gtcccaaccc
     3841 tcgtcgttag aagttgcttc cgaagacgat tttgccatag ccacacgacg cctattaatt
     3901 gtgtcggcta acacgtccgc gatcaaattt gtagttgagc tttttggaat tgcgatcgca
     3961 taacttcgta tagcatacat tatacgaagt tataagctcg gaacgctgcg ctcggtcgtt
     4021 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4081 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4141 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc tgacgagcat cacaaaaatc
     4201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4561 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4741 gatctcaaga agatcctttg ttagaaaaac tcatcgagca tcaagtgaaa ctgcaattta
     4801 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     4861 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     4921 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     4981 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     5041 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc accaaccaaa
     5101 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgcc gttaaaagga
     5161 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     5221 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca
     5281 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     5341 ataaattccg tcagccagtt tagcctgacc atctcatctg taacatcatt ggcaacgcta
     5401 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     5461 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     5521 atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcatactc
     5581 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgtc cgcgcgtt
//
