LOCUS       sequence-180872-        5544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of microtuble plus ends (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5544)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 5544)
  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"; journalName: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: EB3-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..5544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      57..66
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             927..1622
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1060..1078)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1328..1347
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1743..1864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1780..1799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1834..1853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1871..2326)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2008..2029)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2220..2239
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2353..2457
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2459..2816
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2459..2650
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2482..2502)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2667..2802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2851..3645
                     /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             2851..3645
                     /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(2905..2924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3515..3534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3833..3852)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3877..3924
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4253..4841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4742..4761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5016..5319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5320..5523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5474..5494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cgctagcgct accggactca gatctcgagc tcaagcttcg aattctgcag tcgactgcca
       61 ccatggccgt caatgtgtac tccacatctg tgaccagtga aaatctgagt cgccatgata
      121 tgcttgcatg ggtcaacgac tccctgcacc tcaactatac caagatagaa cagctttgtt
      181 caggggcagc ctactgccag ttcatggaca tgctcttccc cggctgtgtg cacttgagga
      241 aagtgaagtt ccaggccaaa ctagagcatg aatacatcca caacttcaag gtgctgcaag
      301 cagctttcaa gaagatgggt gttgacaaaa tcattcctgt agagaaatta gtgaaaggaa
      361 aattccaaga taattttgag tttattcagt ggtttaagaa attctttgac gcaaactatg
      421 atggaaagga ttacaaccct ctgctggcgc ggcagggcca ggacgtagcg ccacctccta
      481 acccaggtga tcagatcttc aacaaatcca agaaactcat tggcacagca gttccacaga
      541 ggacgtcccc cacaggccca aaaaacatgc agacctctgg ccggctgagc aatgtggccc
      601 ccccctgcat tctccggaag aatcctccat cagcccgaaa tggcggccat gagactgatg
      661 cccaaattct tgaactcaac caacagctgg tggacttgaa gctgacagtg gatgggctgg
      721 agaaggaacg tgacttctac ttcagcaaac ttcgtgacat cgagctcatc tgccaggagc
      781 atgaaagtga aaacagccct gttatctcag gcatcattgg catcctctat gccacagagg
      841 aaggattcgc accccctgag gacgatgaga ttgaagagca tcaacaagaa gaccaggacg
      901 agtacctgga tccaccggtc gccaccatgg tgagcaaggg cgaggcagtg atcaaggagt
      961 tcatgcggtt caaggtgcac atggagggct ccatgaacgg ccacgagttc gagatcgagg
     1021 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1081 gtggccccct gcccttctcc tgggacatcc tgtcccctca gttcatgtac ggctccaggg
     1141 ccttcatcaa gcaccccgcc gacatccccg actactataa gcagtccttc cccgagggct
     1201 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg acccaggaca
     1261 cctccctgga ggacggcacc ctgatctaca aggtgaagct ccgcggcacc aacttccctc
     1321 ctgacggccc cgtaatgcag aagaagacaa tgggctggga agcatccacc gagcggttgt
     1381 accccgagga cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg aaggacggcg
     1441 gccgctacct ggcggacttc aagaccacct acaaggccaa gaagcccgtg cagatgcccg
     1501 gcgcctacaa cgtcgaccgc aagttggaca tcacctccca caacgaggac tacaccgtgg
     1561 tggaacagta cgaacgctcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     1621 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1681 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1741 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1861 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1921 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     1981 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2041 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2101 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2161 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2221 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2281 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2341 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2401 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2461 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2521 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2581 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2641 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2701 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2761 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     2821 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2881 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2941 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3001 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3061 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3121 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3181 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3241 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3301 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3361 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3421 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3481 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3541 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3601 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3661 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3721 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3781 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     3841 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3901 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     3961 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4021 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4081 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4141 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4201 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4261 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4321 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4381 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4441 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4501 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4561 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4621 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4681 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4741 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4801 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4861 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4921 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     4981 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     5041 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     5101 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     5161 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5221 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5281 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     5341 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     5401 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     5461 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     5521 gctggtttag tgaaccgtca gatc
//
