LOCUS       sequence-200532-        3929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mKate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3929)
  AUTHORS   Balleza E, Kim JM, Cluzel P
  TITLE     Systematic characterization of maturation time of fluorescent 
            proteins in living cells.
  JOURNAL   Nat Methods. 2018 Jan;15(1):47-51. doi: 10.1038/nmeth.4509. Epub 
            2017 Nov 20.
  PUBMED    29320486
REFERENCE   2  (bases 1 to 3929)
  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/nmeth.4509"; journalName: "Nat Methods"; date: "2018-01"; 
            volume: "15"; issue: "1"; pages: "47-51"
COMMENT     Sequence Label: pEB2-mKate2
FEATURES             Location/Qualifiers
     source          1..3929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..84
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             269..964
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     terminator      1002..1048
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1221..2171)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2219..2441
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2947..2981)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3022..3816)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3133..3152)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3743..3762
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 agggcgcccc agctggcaat tccgacgtct aagaaaccat tattatcatg acattaacct
       61 ataaaaatag gcgtatcacg aggccctttc gtcttcacca cagctaacac cacgtcgtcc
      121 ctatctgctg ccctaggtct atgagtggtt gctggataac tttacgggca tgcataaggc
      181 tcgtatgata tattcaggga gaccacaacg gtttccctct acaaataatt ttgtttaact
      241 tttctagatt taagaaggag atatacatat gtctgagctg attaaggaga acatgcacat
      301 gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga
      361 aggcaagccc tacgagggca cccagaccat gagaatcaag gcggtcgagg gcggcccttt
      421 acccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa
      481 ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga
      541 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcttaca
      601 ggacggctgc ttgatctaca acgtcaagat cagaggggtg aacttcccat ccaacggccc
      661 tgtgatgcag aagaaaacat taggctggga ggcctccacc gagaccctgt accccgctga
      721 cggcggcctg gaaggcagag ccgacatggc cctgaagtta gtgggcgggg gccacctgat
      781 ctgcaacttg aagaccacat acagatccaa gaaacccgct aagaacttga agatgcccgg
      841 cgtctactat gtggacagaa gactggaaag aatcaaggag gccgacaaag agacctacgt
      901 cgagcagcac gaggtggctg tggccagata ctgcgacctg cctagcaaac tggggcacag
      961 ataaatgtcc agacctgcag gcatgcaagc tctagaggca tcaaataaaa cgaaaggctc
     1021 agtcgaaaga ctgggccttt cgttttatct gttgaaatgc accaaaaact cgtaaaagct
     1081 ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg
     1141 atatctaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag
     1201 atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg
     1261 gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatgctt actttgcatg
     1321 tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt
     1381 agtgtttttc ttagtccgtt acgtaggtag gaatctgatg taatggttgt tggtattttg
     1441 tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct
     1501 agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc
     1561 atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt
     1621 ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc
     1681 tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc
     1741 ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt
     1801 tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt
     1861 gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg
     1921 atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt
     1981 tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct
     2041 ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg
     2101 gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact
     2161 aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg
     2221 gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg taaattctgc
     2281 tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct
     2341 ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa
     2401 aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg
     2461 cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac
     2521 cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc
     2581 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
     2641 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
     2701 gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg
     2761 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc
     2821 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta
     2881 aggcagcggt atcatcaaca ggcttacccg tcttactgtc aaccagaccc gccaggattc
     2941 tcatgcataa aaaacgcccg gcggcaaccg agcgttctga attaattaat catcgcgact
     3001 gcagagatcc cagagtcctt attagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     3061 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     3121 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     3181 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     3241 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
     3301 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     3361 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     3421 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     3481 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     3541 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     3601 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     3661 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     3721 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     3781 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     3841 ttgatcagat cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     3901 tactttgcag ggcttcccaa ccttaccag
//
