LOCUS       sequence-223510-        6973 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 6973)
  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 6973)
  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: EB3-3xmScarletI-N1
FEATURES             Location/Qualifiers
     source          1..6973
                     /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..1619
                     /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
                     STGGMDELY"
     CDS             1638..2330
                     /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
                     STGGMDELY"
     CDS             2349..3044
                     /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"
     polyA_signal    3172..3293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3209..3228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3263..3282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3300..3755)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3437..3458)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3649..3668
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3782..3886
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3888..4245
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3888..4079
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3911..3931)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4096..4231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4280..5074
                     /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             4280..5074
                     /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(4334..4353)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4944..4963
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5262..5281)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5306..5353
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5682..6270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6171..6190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6445..6748
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6749..6952
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6903..6923
                     /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 agcgtccacc 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 gagctgtacc
     1621 agggtggcag cggtggcatg gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt
     1681 tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     1741 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     1801 tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg gccttcatca
     1861 agcaccccgc cgacatcccc gactactata agcagtcctt ccccgagggc ttcaagtggg
     1921 agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt gacccaggac acctccctgg
     1981 aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct cctgacggcc
     2041 ccgtaatgca gaagaagaca atgggctggg aagcgtccac cgagcggttg taccccgagg
     2101 acggcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc ggccgctacc
     2161 tggcggactt caagaccacc tacaaggcca agaagcccgt gcagatgccc ggcgcctaca
     2221 acgtcgaccg caagttggac atcacctccc acaacgagga ctacaccgtg gtggaacagt
     2281 acgaacgctc cgagggccgc cactccaccg gcggcatgga cgagctgtac cagggtggca
     2341 gcggtggcat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     2401 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2461 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     2521 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg
     2581 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     2641 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     2701 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     2761 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     2821 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     2881 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     2941 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     3001 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagaa ttggccgcga
     3061 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3121 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3181 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3241 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3301 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3361 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3421 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3481 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3541 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3601 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3661 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3721 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3781 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3841 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3901 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3961 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4021 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4081 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4141 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4201 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4261 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4321 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4381 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4441 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4501 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4561 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4621 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4681 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4741 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4801 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4861 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4921 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4981 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5041 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5101 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5161 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5221 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5281 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5341 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5401 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5461 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5521 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6361 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     6421 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6481 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     6541 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     6601 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6661 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6721 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     6781 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     6841 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     6901 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     6961 gaaccgtcag atc
//
