LOCUS       sequence-200521-        3944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mRuby3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3944)
  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 3944)
  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-mRuby3
FEATURES             Location/Qualifiers
     source          1..3944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..36
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      969..1015
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1188..2138)
                     /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      2186..2408
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2914..2948)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2989..3783)
                     /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(3100..3119)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3710..3729
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac cacagctaac
       61 accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata actttacggg
      121 catgcataag gctcgtatga tatattcagg gagaccacaa cggtttccct ctacaaataa
      181 ttttgtttaa cttttctaga tttaagaagg agatatacat atgagtaaag gagaagaact
      241 gatcaaggaa aatatgcgta tgaaggtggt catggaaggt tcggtcaacg gccaccaatt
      301 caaatgcaca ggtgaaggag aaggcagacc gtacgaggga gtgcaagtga tgaggatcaa
      361 agtcatcgag ggaggacccc tgccatttgc ctttgacatt cttgccacgt cgttcatgta
      421 tggcagccgt acctttatca agtacccggc cgacatccct gatttcttta aacagtcctt
      481 tcctgagggt tttacttggg aaagagttac gagatacgaa gatggtggag tcgtcaccgt
      541 cacgcaggac accagccttg aggatggcga gctcgtctac aacgtcaagg tcagaggggt
      601 aaactttccc tccaatggtc ccgtgatgca gaagaagacc aagggttggg agcctaatac
      661 agagatgatg tatccagcag atggtggtct gagaggatac actgacatcg cactgaaagt
      721 tgatggtggt ggccatctgc actgcaactt cgtgacaact tacaggtcaa aaaagaccgt
      781 cgggaacatc aagatgcccg gtgtccatgc cgttgatcac cgcctggaaa ggatcgagga
      841 gagtgacaat gaaacctacg tagtgcaaag agaagtggca gttgccaaat acagcaacct
      901 tggtggtggc atggatgaac tatacaaata aatgtccaga cctgcaggca tgcaagctct
      961 agaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1021 gaaatgcacc aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat
     1081 ctgtgcatat ggacagtttt ccctttgata tctaacggtg aacagttgtt ctacttttgt
     1141 ttgttagtct tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg
     1201 tatttagcca gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa
     1261 ccattgagat catgcttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc
     1321 tgaatttttg cagttaaagc atcgtgtagt gtttttctta gtccgttacg taggtaggaa
     1381 tctgatgtaa tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt
     1441 tcggttacga gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg
     1501 cggcctcgct tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt
     1561 ggtttcaaaa cccattggtt aagcctttta aactcatggt agttattttc aagcattaac
     1621 atgaacttaa attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt
     1681 agttctttta ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca
     1741 tgttccagat tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca
     1801 ctaaaaacta attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc
     1861 tcaaagcctt taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt
     1921 gctttagcta atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg
     1981 ttataagtga acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt
     2041 agtgccacac agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc
     2101 gctagttcat ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga
     2161 ttttaatcac tataccaatt gagatgggct agtcaatgat aattactagt ccttttcctt
     2221 tgagttgtgg gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct
     2281 agaccctctg taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt
     2341 ataatttata gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg
     2401 tataactcac tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt
     2461 gctcctctac aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg
     2521 ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt
     2581 tcgtgacatt cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta
     2641 caggcgcctt ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac
     2701 gggcttctca gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt
     2761 tcagcagttc ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt
     2821 cattcagact ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct
     2881 tactgtcaac cagacccgcc aggattctca tgcataaaaa acgcccggcg gcaaccgagc
     2941 gttctgaatt aattaatcat cgcgactgca gagatcccag agtccttatt agaagaactc
     3001 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     3061 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     3121 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     3181 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     3241 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     3301 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     3361 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     3421 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     3481 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     3541 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     3601 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     3661 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     3721 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     3781 catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat
     3841 ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
     3901 cgccccagct ggcaattccg acgtctaaga aaccattatt atca
//
