LOCUS       Exported                5563 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-mNeonGreen
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5563)
  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 5563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5563
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        105..462
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(128..148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      313..448
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     375..394
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             497..1291
                     /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(551..570)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1161..1180
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1479..1498)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1523..1570
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1899..2487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2388..2407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2662..2965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2966..3169
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3120..3140
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3247..3256
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      4111..4120
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4117..4827
                     /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      4372..4381
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    4952..5073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4989..5008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5043..5062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5080..5535)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5217..5238)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5429..5448
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5562..103
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
       61 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
      121 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
      181 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
      241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
      301 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
      361 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
      421 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
      481 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
      541 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
      601 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
      661 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
      721 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
      781 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
      841 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
      901 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
      961 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     1021 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     1081 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     1141 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     1201 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     1261 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     1321 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     1381 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     1441 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     1501 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     1561 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     1621 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     1681 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     1741 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1801 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1861 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1921 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1981 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2041 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2101 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2161 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2221 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2281 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2341 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2401 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2461 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2521 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2581 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     2641 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2701 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2761 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2821 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2881 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2941 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     3001 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3061 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     3121 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     3181 ccgtcagatc cgctagcgct accggactca gatctcgagc tcaagcttcg aattctgcag
     3241 tcgactgcca ccatggccgt caatgtgtac tccacatctg tgaccagtga aaatctgagt
     3301 cgccatgata tgcttgcatg ggtcaacgac tccctgcacc tcaactatac caagatagaa
     3361 cagctttgtt caggggcagc ctactgccag ttcatggaca tgctcttccc cggctgtgtg
     3421 cacttgagga aagtgaagtt ccaggccaaa ctagagcatg aatacatcca caacttcaag
     3481 gtgctgcaag cagctttcaa gaagatgggt gttgacaaaa tcattcctgt agagaaatta
     3541 gtgaaaggaa aattccaaga taattttgag tttattcagt ggtttaagaa attctttgac
     3601 gcaaactatg atggaaagga ttacaaccct ctgctggcgc ggcagggcca ggacgtagcg
     3661 ccacctccta acccaggtga tcagatcttc aacaaatcca agaaactcat tggcacagca
     3721 gttccacaga ggacgtcccc cacaggccca aaaaacatgc agacctctgg ccggctgagc
     3781 aatgtggccc ccccctgcat tctccggaag aatcctccat cagcccgaaa tggcggccat
     3841 gagactgatg cccaaattct tgaactcaac caacagctgg tggacttgaa gctgacagtg
     3901 gatgggctgg agaaggaacg tgacttctac ttcagcaaac ttcgtgacat cgagctcatc
     3961 tgccaggagc atgaaagtga aaacagccct gttatctcag gcatcattgg catcctctat
     4021 gccacagagg aaggattcgc accccctgag gacgatgaga ttgaagagca tcaacaagaa
     4081 gaccaggacg agtacctgga tccaccggtc gccaccatgg tgagcaaggg cgaggaggat
     4141 aacatggcct ctctcccagc gacacatgag ttacacatct ttggctccat caacggtgtg
     4201 gactttgaca tggtgggtca gggcaccggc aatccaaatg atggttatga ggagttaaac
     4261 ctgaagtcca ccaagggtga cctccagttc tccccctgga ttctggtccc tcatatcggg
     4321 tatggcttcc atcagtacct gccctaccct gacgggatgt cgcctttcca ggccgccatg
     4381 gtagatggct ccggctacca agtccatcgc acaatgcagt ttgaagatgg tgcctccctt
     4441 actgttaact accgctacac ctacgaggga agccacatca aaggagaggc ccaggtgaag
     4501 gggactggtt tccctgctga cggtcctgtg atgaccaact cgctgaccgc tgcggactgg
     4561 tgcaggtcga agaagactta ccccaacgac aaaaccatca tcagtacctt taagtggagt
     4621 tacaccactg gaaatggcaa gcgctaccgg agcactgcgc ggaccaccta cacctttgcc
     4681 aagccaatgg cggctaacta tctgaagaac cagccgatgt acgtgttccg taagacggag
     4741 ctcaagcact ccaagaccga gctcaacttc aaggagtggc aaaaggcctt taccgatgtg
     4801 atgggcatgg acgagctgta caagtaagaa ttggccgcga ctctagatca taatcagcca
     4861 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4921 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4981 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5041 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     5101 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5161 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5221 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5281 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5341 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5401 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5461 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5521 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcg
//
