LOCUS       Exported                4784 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of the nucleus (can be used for colocalization
            studies).
ACCESSION   .
VERSION     .
KEYWORDS    3xnls-mNeonGreen
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4784)
  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 4784)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4784
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     200..220
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(245..349)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      376..831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(463..482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     673..694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    838..959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(849..868)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     903..922
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1082..1792)
                     /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      1528..1537
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      1789..1798
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(1811..1831)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1835..1855)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1859..1879)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1963..2166)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1992..2012)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2167..2470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2645..3233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2725..2744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3562..3609
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3634..3653
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3841..4635)
                     /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(3952..3971)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4562..4581
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4670..243)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4684..35
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4738..4757)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
       61 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      121 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
      181 gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct
      241 caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      301 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      361 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      421 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      481 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      541 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
      601 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
      661 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
      721 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
      781 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tacgccttaa
      841 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      901 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      961 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1021 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatctagag tcgcggccgc
     1081 tttacttgta cagctcgtcc atgcccatca catcggtaaa ggccttttgc cactccttga
     1141 agttgagctc ggtcttggag tgcttgagct ccgtcttacg gaacacgtac atcggctggt
     1201 tcttcagata gttagccgcc attggcttgg caaaggtgta ggtggtccgc gcagtgctcc
     1261 ggtagcgctt gccatttcca gtggtgtaac tccacttaaa ggtactgatg atggttttgt
     1321 cgttggggta agtcttcttc gacctgcacc agtccgcagc ggtcagcgag ttggtcatca
     1381 caggaccgtc agcagggaaa ccagtcccct tcacctgggc ctctcctttg atgtggcttc
     1441 cctcgtaggt gtagcggtag ttaacagtaa gggaggcacc atcttcaaac tgcattgtgc
     1501 gatggacttg gtagccggag ccatctacca tggcggcctg gaaaggcgac atcccgtcag
     1561 ggtagggcag gtactgatgg aagccatacc cgatatgagg gaccagaatc cagggggaga
     1621 actggaggtc acccttggtg gacttcaggt ttaactcctc ataaccatca tttggattgc
     1681 cggtgccctg acccaccatg tcaaagtcca caccgttgat ggagccaaag atgtgtaact
     1741 catgtgtcgc tgggagagag gccatgttat cctcctcgcc cttgctcacc atggtggcga
     1801 ccggtgatcc cactttccgt ttctttttgg gatcgacctt acgctttttc ttcgggtcta
     1861 cctttctctt cttttttgga tctgatccca tggtgggaat tcgaagcttg agctcgagat
     1921 ctgagtccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc ttatatagac
     1981 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2041 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2101 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     2161 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     2221 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     2281 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     2341 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2401 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2461 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2521 ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3061 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3361 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga
     3421 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag
     3481 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct
     3541 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt
     3601 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     3661 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     3721 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     3781 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     3841 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     3901 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     3961 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     4021 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     4081 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc
     4141 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     4201 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     4261 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     4321 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     4381 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     4441 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     4501 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     4561 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     4621 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc
     4681 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     4741 cggcctctgc ataaataaaa aaaattagtc agccatgggg cgga
//
