LOCUS       Exported                7009 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of microtuble plus ends (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    EB3-3xmNeonGreen-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7009)
  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 160374
REFERENCE   2  (bases 1 to 7009)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7009
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      57..66
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      921..930
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             927..1631
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      1182..1191
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1650..2354
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      1905..1914
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2373..3083
                     /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      2628..2637
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    3208..3329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3245..3264)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3299..3318
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3336..3791)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3473..3494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3685..3704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3818..3922
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3924..4281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3947..3967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4132..4267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4194..4213
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4316..5110
                     /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(4370..4389)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4980..4999
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5298..5317)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5342..5389
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5718..6306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6207..6226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6481..6784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6785..6988
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6939..6959
                     /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 cgaggaggat aacatggcct
      961 ctctcccagc gacacatgag ttacacatct ttggctccat caacggtgtg gactttgaca
     1021 tggtgggtca gggcaccggc aatccaaatg atggttatga ggagttaaac ctgaagtcca
     1081 ccaagggtga cctccagttc tccccctgga ttctggtccc tcatatcggg tatggcttcc
     1141 atcagtacct gccctaccct gacgggatgt cgcctttcca ggccgccatg gtagatggct
     1201 ccggctacca agtccatcgc acaatgcagt ttgaagatgg tgcctccctt actgttaact
     1261 accgctacac ctacgaggga agccacatca aaggagaggc ccaggtgaag gggactggtt
     1321 tccctgctga cggtcctgtg atgaccaact cgctgaccgc tgcggactgg tgcaggtcga
     1381 agaagactta ccccaacgac aaaaccatca tcagtacctt taagtggagt tacaccactg
     1441 gaaatggcaa gcgctaccgg agcactgcgc ggaccaccta cacctttgcc aagccaatgg
     1501 cggctaacta tctgaagaac cagccgatgt acgtgttccg taagacggag ctcaagcact
     1561 ccaagaccga gctcaacttc aaggagtggc aaaaggcctt taccgatgtg atgggcatgg
     1621 acgagctgta ccagggtggc agcggtggca tggtgagcaa gggcgaggag gataacatgg
     1681 cctctctccc agcgacacat gagttacaca tctttggctc catcaacggt gtggactttg
     1741 acatggtggg tcagggcacc ggcaatccaa atgatggtta tgaggagtta aacctgaagt
     1801 ccaccaaggg tgacctccag ttctccccct ggattctggt ccctcatatc gggtatggct
     1861 tccatcagta cctgccctac cctgacggga tgtcgccttt ccaggccgcc atggtagatg
     1921 gctccggcta ccaagtccat cgcacaatgc agtttgaaga tggtgcctcc cttactgtta
     1981 actaccgcta cacctacgag ggaagccaca tcaaaggaga ggcccaggtg aaggggactg
     2041 gtttccctgc tgacggtcct gtgatgacca actcgctgac cgctgcggac tggtgcaggt
     2101 cgaagaagac ttaccccaac gacaaaacca tcatcagtac ctttaagtgg agttacacca
     2161 ctggaaatgg caagcgctac cggagcactg cgcggaccac ctacaccttt gccaagccaa
     2221 tggcggctaa ctatctgaag aaccagccga tgtacgtgtt ccgtaagacg gagctcaagc
     2281 actccaagac cgagctcaac ttcaaggagt ggcaaaaggc ctttaccgat gtgatgggca
     2341 tggacgagct gtaccagggt ggcagcggtg gcatggtgag caagggcgag gaggataaca
     2401 tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac ggtgtggact
     2461 ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag ttaaacctga
     2521 agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat atcgggtatg
     2581 gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc gccatggtag
     2641 atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc tcccttactg
     2701 ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag gtgaagggga
     2761 ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg gactggtgca
     2821 ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag tggagttaca
     2881 ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc tttgccaagc
     2941 caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag acggagctca
     3001 agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg
     3061 gcatggacga gctgtacaag taagaattgg ccgcgactct agatcataat cagccatacc
     3121 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3181 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3301 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3361 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3421 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3481 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3541 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3601 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3661 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3721 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3781 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3901 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3961 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4141 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4201 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4261 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4321 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4381 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4441 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4501 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4561 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4621 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4681 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4741 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4801 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4861 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4921 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4981 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5041 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5101 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5161 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5221 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5281 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5341 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5401 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5461 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5521 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5581 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5641 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5701 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5761 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5821 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5881 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5941 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6001 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6061 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6121 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6181 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6241 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6301 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6361 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     6421 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     6481 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     6541 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     6601 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     6661 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     6721 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     6781 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     6841 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     6901 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     6961 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatc
//
