LOCUS       sequence-376485-        5554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for labelling microtubule plus ends in mammalian cells 
            with EB3-mScarlet3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  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 5554)
  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: pEB3-mScarlet3_N1
FEATURES             Location/Qualifiers
     source          1..5554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             169..963
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             169..963
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(223..242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     833..852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1151..1170)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1195..1242
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1571..2159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2060..2079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2334..2637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2638..2841
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2792..2812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2919..2928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3783..3792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3789..4475
                     /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     4187..4206
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4615..4736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4652..4671)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4706..4725
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4743..5198)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4880..4901)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5092..5111
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5225..5329
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5331..5554,1..134)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5331..5522
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5354..5374)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5539..5554,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
       61 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      121 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      181 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      241 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      301 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
      361 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      421 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      481 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      541 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      601 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      661 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
      721 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      781 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      841 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
      901 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
      961 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     1021 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1081 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     1141 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     1201 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     1261 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     1321 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     1381 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     1441 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1501 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1681 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2221 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     2281 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2341 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2401 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2461 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2521 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2581 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2641 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2701 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2761 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2821 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
     2881 cagatctcga gctcaagctt cgaattctgc agtcgactgc caccatggcc gtcaatgtgt
     2941 actccacatc tgtgaccagt gaaaatctga gtcgccatga tatgcttgca tgggtcaacg
     3001 actccctgca cctcaactat accaagatag aacagctttg ttcaggggca gcctactgcc
     3061 agttcatgga catgctcttc cccggctgtg tgcacttgag gaaagtgaag ttccaggcca
     3121 aactagagca tgaatacatc cacaacttca aggtgctgca agcagctttc aagaagatgg
     3181 gtgttgacaa aatcattcct gtagagaaat tagtgaaagg aaaattccaa gataattttg
     3241 agtttattca gtggtttaag aaattctttg acgcaaacta tgatggaaag gattacaacc
     3301 ctctgctggc gcggcagggc caggacgtag cgccacctcc taacccaggt gatcagatct
     3361 tcaacaaatc caagaaactc attggcacag cagttccaca gaggacgtcc cccacaggcc
     3421 caaaaaacat gcagacctct ggccggctga gcaatgtggc ccccccctgc attctccgga
     3481 agaatcctcc atcagcccga aatggcggcc atgagactga tgcccaaatt cttgaactca
     3541 accaacagct ggtggacttg aagctgacag tggatgggct ggagaaggaa cgtgacttct
     3601 acttcagcaa acttcgtgac atcgagctca tctgccagga gcatgaaagt gaaaacagcc
     3661 ctgttatctc aggcatcatt ggcatcctct atgccacaga ggaaggattc gcaccccctg
     3721 aggacgatga gattgaagag catcaacaag aagaccagga cgagtacctg gatccaccgg
     3781 tcgccaccat ggatagcacc gaggcagtga tcaaggagtt catgcggttc aaggtgcaca
     3841 tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3901 acgagggcac ccagaccgcc aagctgaggg tgaccaaggg tggccccctg cccttctcct
     3961 gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcacgaag caccccgccg
     4021 acatccccga ctactggaag cagtccttcc ccgagggctt caagtgggag cgcgtgatga
     4081 acttcgagga cggcggcgcc gtgtccgtgg cccaggacac ctccctggag gacggcaccc
     4141 tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc gtaatgcaga
     4201 agaagacaat gggctgggaa gcatccaccg agcggttgta ccccgaggac gtcgtgctga
     4261 agggcgacat taagatggcc ctgcgcctga aggacggcgg ccgctacctg gcggacttca
     4321 agaccaccta cagggccaag aagcccgtgc agatgcccgg cgccttcaac atcgaccgca
     4381 agttggacat cacatcccac aacgaggact acaccgtggt ggaacagtac gaacgctccg
     4441 tggcccgcca ctccaccggc ggctccggtg gctcctaact gtacaagtaa gaattggccg
     4501 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4561 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4621 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4681 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     4741 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     4801 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     4861 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4921 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4981 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5041 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5101 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5161 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     5221 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5281 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     5341 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5401 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5461 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5521 ccgcccctaa ctccgcccat cccgccccta actc
//
