LOCUS       sequence-384158-        5548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mNeonGreen fused to EB3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5548)
  AUTHORS   Schneider MWG, Gibson BA, Otsuka S, Spicer MFD, Petrovic M, Blaukopf
            C, Langer CCH, Batty P, Nagaraju T, Doolittle LK, Rosen MK, Gerlich 
            DW
  TITLE     A mitotic chromatin phase transition prevents perforation by 
            microtubules.
  JOURNAL   Nature. 2022 Sep;609(7925):183-190. doi: 10.1038/s41586-022-05027-y.
            Epub 2022 Aug 3.
  PUBMED    35922507
REFERENCE   2  (bases 1 to 5548)
  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/s41586-022-05027-y"; journalName: "Nature"; date: 
            "2022-09"; volume: "609"; issue: "7925"; pages: "183-190"
COMMENT     Sequence Label: EB3-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..5548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        159..462
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        463..666
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     617..637
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      744..753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1602..2312
                     /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      1857..1866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2434..2555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2471..2490)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2525..2544
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2562..3017)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2699..2720)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2911..2930
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3044..3148
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3150..3507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3150..3341
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3173..3193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3358..3493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3542..4336
                     /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             3542..4336
                     /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(3596..3615)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4206..4225
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4524..4543)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4568..4615
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4944..5532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5433..5452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
       61 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
      121 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      181 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      241 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      301 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      361 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      421 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      481 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      541 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      601 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      661 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca
      721 agcttcgaat tctgcagtcg actgccacca tggccgtcaa tgtgtactcc acatctgtga
      781 ccagtgaaaa tctgagtcgc catgatatgc ttgcatgggt caacgactcc ctgcacctca
      841 actataccaa gatagaacag ctttgttcag gggcagccta ctgccagttc atggacatgc
      901 tcttccccgg ctgtgtgcac ttgaggaaag tgaagttcca ggccaaacta gagcatgaat
      961 acatccacaa cttcaaggtg ctgcaagcag ctttcaagaa gatgggtgtt gacaaaatca
     1021 ttcctgtaga gaaattagtg aaaggaaaat tccaagataa ttttgagttt attcagtggt
     1081 ttaagaaatt ctttgacgca aactatgatg gaaaggatta caaccctctg ctggcgcggc
     1141 agggccagga cgtagcgcca cctcctaacc caggtgatca gatcttcaac aaatccaaga
     1201 aactcattgg cacagcagtt ccacagagga cgtcccccac aggcccaaaa aacatgcaga
     1261 cctctggccg gctgagcaat gtggcccccc cctgcattct ccggaagaat cctccatcag
     1321 cccgaaatgg cggccatgag actgatgccc aaattcttga actcaaccaa cagctggtgg
     1381 acttgaagct gacagtggat gggctggaga aggaacgtga cttctacttc agcaaacttc
     1441 gtgacatcga gctcatctgc caggagcatg aaagtgaaaa cagccctgtt atctcaggca
     1501 tcattggcat cctctatgcc acagaggaag gattcgcacc ccctgaggac gatgagattg
     1561 aagagcatca acaagaagac caggacgagt acctggatcc tatggtgagc aagggcgagg
     1621 aggataacat ggcctctctc ccagcgacac atgagttaca catctttggc tccatcaacg
     1681 gtgtggactt tgacatggtg ggtcagggca ccggcaatcc aaatgatggt tatgaggagt
     1741 taaacctgaa gtccaccaag ggtgacctcc agttctcccc ctggattctg gtccctcata
     1801 tcgggtatgg cttccatcag tacctgccct accctgacgg gatgtcgcct ttccaggccg
     1861 ccatggtaga tggctccgga taccaagtcc atcgcacaat gcagtttgaa gatggtgcct
     1921 cccttactgt taactaccgc tacacctacg agggaagcca catcaaagga gaggcccagg
     1981 tgaaggggac tggtttccct gctgacggtc ctgtgatgac caactcgctg accgctgcgg
     2041 actggtgcag gtcgaagaag acttacccca acgacaaaac catcatcagt acctttaagt
     2101 ggagttacac cactggaaat ggcaagcgct accggagcac tgcgcggacc acctacacct
     2161 ttgccaagcc aatggcggct aactatctga agaaccagcc gatgtacgtg ttccgtaaga
     2221 cggagctcaa gcactccaag accgagctca acttcaagga gtggcaaaag gcctttaccg
     2281 atgtgatggg catggacgag ctgtacaagt aagcggccgc gactctagat cataatcagc
     2341 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2401 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2461 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2521 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2581 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2641 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2701 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2761 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2821 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2881 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2941 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3001 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3061 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3121 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3181 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3241 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3301 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3361 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3421 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3481 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3541 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3601 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3661 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3721 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3781 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3841 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3901 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3961 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4021 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4081 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4141 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4201 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4261 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4321 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4381 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4441 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4501 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4561 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4621 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4681 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4741 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4801 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4861 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4921 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4981 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5041 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5101 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5161 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5221 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5281 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5341 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5401 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5461 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5521 agcctatgga aaaacgccag caacgcgg
//
