LOCUS       sequence-273204-        5544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of microtubules.
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: EMTB-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      271..280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             280..309
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1129..1138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1135..1845
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1390..1399
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1968..2089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2005..2024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2059..2078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2096..2551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2233..2254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2445..2464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2578..2682
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2684..3041
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2684..2875
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2707..2727)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2892..3027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3076..3870
                     /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             3076..3870
                     /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(3130..3149)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3740..3759
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4058..4077)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4102..4149
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4478..5066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4967..4986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5241..5544
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc gccaccatgg agcagaagct catctcagaa
      301 gaagacctcg gcgcgccagt gcgaagcgaa acagcacccg acagctacaa agtgcaagat
      361 aagaaaaatg cctccagccg ccctgcctct gcaatttcag gacaaaataa caaccactca
      421 ggaaataaac cagaccctcc gcctgtgtta cgtgttgatg accggcagcg gctggcccgg
      481 gagcgacgtg aggaacggga gaaacagcta gctgcaagag aaatagtgtg gttagaaaga
      541 gaagagcgag ccaggcagca ctacgagaag cacctggaag agcggaagaa gaggttggag
      601 gagcagaggc agaaggagga gcggaggagg gctgctgtgg aggagaagcg gaggcagaga
      661 cttgaggagg acaaagaacg ccacgaagct gttgtacggc gcacaatgga aaggagccag
      721 aagccaaaac agaagcataa ccgttggtcg tggggaggct ctctccatgg gagccctagc
      781 atccacagtg cagatccaga caggcggtca gtttccacca tgaatctttc gaaatatgtt
      841 gatcccgtca ttagcaagcg gctctcctct tcatctgcaa ctttactaaa ttctccagat
      901 agagctcgcc gcctgcagct cagcccatgg gagagcagcg ttgttaacag actcctgacg
      961 cccacacatt cgttcctggc cagaagtaaa agcacagctg ccttgtctgg agaagcagca
     1021 tcttgcagcc ccatcatcat gccctacaaa gctgcacact ctagaaattc gatggatcga
     1081 ccaaaactct ttgtaacacc acctgagggc tcttcggatc caccggtcgc caccatggtg
     1141 agcaagggcg aggaggataa catggcctct ctcccagcga cacatgagtt acacatcttt
     1201 ggctccatca acggtgtgga ctttgacatg gtgggtcagg gcaccggcaa tccaaatgat
     1261 ggttatgagg agttaaacct gaagtccacc aagggtgacc tccagttctc cccctggatt
     1321 ctggtccctc atatcgggta tggcttccat cagtacctgc cctaccctga cgggatgtcg
     1381 cctttccagg ccgccatggt agatggctcc ggctaccaag tccatcgcac aatgcagttt
     1441 gaagatggtg cctcccttac tgttaactac cgctacacct acgagggaag ccacatcaaa
     1501 ggagaggccc aggtgaaggg gactggtttc cctgctgacg gtcctgtgat gaccaactcg
     1561 ctgaccgctg cggactggtg caggtcgaag aagacttacc ccaacgacaa aaccatcatc
     1621 agtaccttta agtggagtta caccactgga aatggcaagc gctaccggag cactgcgcgg
     1681 accacctaca cctttgccaa gccaatggcg gctaactatc tgaagaacca gccgatgtac
     1741 gtgttccgta agacggagct caagcactcc aagaccgagc tcaacttcaa ggagtggcaa
     1801 aaggccttta ccgatgtgat gggcatggac gagctgtaca agtaaagcgg ccgcgactct
     1861 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1921 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     2101 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2161 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2221 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2281 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2341 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2401 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2461 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2521 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2701 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2761 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2941 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3001 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3061 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3121 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3181 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3241 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3301 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3361 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3421 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3481 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3541 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3601 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3661 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3721 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3781 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3841 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3901 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3961 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4021 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4081 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4141 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4201 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4261 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4321 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4381 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4501 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4561 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4621 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4681 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4741 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4801 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4861 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4921 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4981 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5041 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5101 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5161 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     5221 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     5281 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     5341 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     5401 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     5461 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     5521 gtattagtca tcgctattac catg
//
