LOCUS       sequence-405565-        3460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a centromere sequence that is recognized 
            in diatoms with AC overhangs used to build level 1 constructs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3460)
  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 3460)
  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_AC-CEN_ARS_HIS
FEATURES             Location/Qualifiers
     source          1..3460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      829..1417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1318..1337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1586..2248)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(2249..2435)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    2436..2939
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             join(3407..3460,1..738)
                     /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 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
       61 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
      121 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
      181 gagagtgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
      241 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
      301 gctggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
      361 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
      421 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
      481 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
      541 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
      601 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
      661 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
      721 aaggtagtcg gcaaataata atcacctaga atgcatgacc aaaatccctt aacgtgagtt
      781 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      841 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      901 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      961 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1021 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1081 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1261 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1321 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1381 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1441 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1501 ttctgtgggt ggaagggctc ggagttgtgg taatctatgt atcctggtgg tctcacattg
     1561 gtcaagtcca gactcctgtg taaaactaca taagaacacc tttggtggag ggaacatcgt
     1621 tggtaccatt gggcgaggtg gcttctctta tggcaaccgc aagagccttg aacgcactct
     1681 cactacggtg atgatcattc ttgcctcgca gacaatcaac gtggagggta attctgctag
     1741 cctctgcaaa gctttcaaga aaatgcggga tcatctcgca agagagatct cctactttct
     1801 ccctttgcaa accaagttcg acaactgcgt acggcctgtt cgaaagatct accaccgctc
     1861 tggaaagtgc ctcatccaaa ggcgcaaatc ctgatccaaa cctttttact ccacgcacgg
     1921 cccctagggc ctctttaaaa gcttgaccga gagcaatccc gcagtcttca gtggtgtgat
     1981 ggtcgtctat gtgtaagtca ccaatgcact caacgattag cgaccagccg gaatgcttgg
     2041 ccagagcatg tatcatatgg tccagaaacc ctatacctgt gtggacgtta atcacttgcg
     2101 attgtgtggc ctgttctgct actgcttctg cctctttttc tgggaagatc gagtgctcta
     2161 tcgctagggg accacccttt aaagagatcg caatctgaat cttggtttca tttgtaatac
     2221 gctttactag ggctttctgc tctgtcatct ttgccttcgt ttatcttgcc tgctcatttt
     2281 ttagtatatt cttcgaagaa atcacattac tttatataat gtataattca ttatgtgata
     2341 atgccaatcg ctaagaaaaa aaaagagtca tccgctaggt ggaaaaaaaa aatgaaaatc
     2401 attaccgagg cataaaaaaa tatagagtgt actaggatcg cttgcctgta acttacacgc
     2461 gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta
     2521 tgttttgtat ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga
     2581 aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt
     2641 attagacaag aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa
     2701 aatcacagga ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa
     2761 tacctgagag caggacgagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc
     2821 ttttacatct tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt
     2881 tttaatttat atatttatat taaaaaattt aaattataat tatttttata gcacgtgatg
     2941 ctcgctgaag acgagctagc tcctgagacc tgagacgaga cgagacagcc tgagaatgga
     3001 tgcgagtaat gagctcggca caaaatcacc actcgataca ggcagcccat cagtcctgga
     3061 cggcgtcagc gggagagccg ttgtaaggcg gcagactttg ctcatgttac cgatgctatt
     3121 cggaagaacg gcaactaagc tgccgggttt gaaacacgga tgatctcgcg gagggtagca
     3181 tgttgattgt aacgatgaca gagcgttgct gcctgtgatc accgctcctc tagagtcgac
     3241 ttgcaggcat gcaagcttag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
     3301 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     3361 caggggatca atatctgatc aagagacagg atgaggatcg tttcgcatga gggaagcggt
     3421 gatcgccgaa gtatcgactc aactatcaga ggtagttggc
//
