LOCUS       sequence-405615-        2771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a mRuby3 fluorescent protein with DE 
            overhangs used to build a level 1 construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2771)
  AUTHORS   Pollak B, Matute T, Nunez I, Cerda A, Lopez C, Vargas V, Kan A, 
            Bielinski V, von Dassow P, Dupont CL, Federici F
  TITLE     Universal loop assembly: open, efficient and cross-kingdom DNA 
            fabrication.
  JOURNAL   Synth Biol (Oxf). 2020;5(1):ysaa001. doi: 10.1093/synbio/ysaa001. 
            Epub 2020 Feb 5.
  PUBMED    32161816
REFERENCE   2  (bases 1 to 2771)
  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"; journalName: "Synth Biol 
            (Oxf)."; date: "2020"; volume: "5(1):ysaa001. doi"; pages: " 
            10.1093/synbio/ysaa001. Epub 2020 Feb 5"
COMMENT     Sequence Label: pL0_DE-mRuby3
FEATURES             Location/Qualifiers
     source          1..2771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      843..1431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1332..1351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1577..2284)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     CDS             join(2732..2771,1..752)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
       61 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
      121 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
      181 cggcttcccc tggagagagt gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
      241 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
      301 gcaatgacat tcttgctggt atcttcgagc cagccacgat cgacattgat ctggctatct
      361 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
      421 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
      481 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
      541 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
      601 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
      661 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
      721 aaggcgagat caccaaggta gtcggcaaat aataatcacc tagaatgcat gaccaaaatc
      781 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      841 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      901 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      961 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1021 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1081 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1141 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1201 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1261 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1321 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1381 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1441 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     1501 gcgttatccc ctgattctgt gggtggaagg gctcggagtt gtggtaatct atgtatcctg
     1561 gtggtctcta agcttacttg tacagctcgt ccatgccacc accaaggttg ctgtatttgg
     1621 caactgccac ttctctttgc actacgtagg tttcattgtc actctcctcg atcctttcca
     1681 ggcggtgatc aacggcatgg acaccgggca tcttgatgtt cccgacggtc ttttttgacc
     1741 tgtaagttgt cacgaagttg cagtgcagat ggccaccacc atcaactttc agtgcgatgt
     1801 cagtgtatcc tctcagacca ccatctgctg gatacatcat ctctgtatta ggctcccaac
     1861 ccttggtctt cttctgcatc acgggaccat tggagggaaa gtttacccct ctgaccttga
     1921 cgttgtagac gagctcgcca tcctcaaggc tggtgtcctg cgtgacggtg acgactccac
     1981 catcttcgta tctcgtaact ctttcccaag taaaaccctc aggaaaggac tgtttaaaga
     2041 aatcagggat gtcggccggg tacttgataa aggtacggct gccatacatg aacgacgtgg
     2101 caagaatgtc aaaggcaaat ggcaggggtc ctccctcgat gactttgatc ctcatggttt
     2161 gcactccctc gtacggtctg ccttctcctt cacctgtgca tttgaattgg tggccgttga
     2221 ccgaaccttc catgaccacc ttcatacgca tattttcctt gatcagttct tcgcccttag
     2281 acacaccttg agacctgaga cgagacgaga cagcctgaga atggatgcga gtaatgagct
     2341 cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag
     2401 agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac
     2461 taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga
     2521 tgacagagcg ttgctgcctg tgatcaccgc tcctctagag tcgacttgca ggcatgcaag
     2581 cttagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     2641 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaatatc
     2701 tgatcaagag acaggatgag gatcgtttcg catgagggaa gcggtgatcg ccgaagtatc
     2761 gactcaacta t
//
