LOCUS       sequence-405695-        3487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a phleomycin antibiotic cassette (fcpF 
            promoter) with EF overhangs used to build a level 1 construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3487)
  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 3487)
  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_EF-PtBle
FEATURES             Location/Qualifiers
     source          1..3487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             406..1197
                     /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"
     rep_origin      1288..1876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1777..1796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2327..2701)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(3405..3423)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gcgagtaatg agctcggcac aaaatcacca ctcgatacag gcagcccatc agtccgggac
       61 ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc gatgctattc
      121 ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg agggtagcat
      181 gttgattgta acgatgacag agcgttgctg cctgtgatca ccgctcctct agagtcgact
      241 tgcaggcatg caagcttagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
      301 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
      361 aggggatcaa tatctgatca agagacagga tgaggatcgt ttcgcatgag ggaagcggtg
      421 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
      481 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
      541 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
      601 ttgatcaacg accttttgga aacttcggct tcccctggag agagtgagat tctccgcgct
      661 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
      721 gaactgcaat ttggagaatg gcagcgcaat gacattcttg ctggtatctt cgagccagcc
      781 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
      841 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
      901 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
      961 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     1021 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     1081 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     1141 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataataa
     1201 tcacctagaa tgcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1261 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1321 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1381 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1441 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1501 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1561 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1621 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1681 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1741 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1801 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1861 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1921 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtgggtg gaagggctcg
     1981 gagttgtggt aatctatgta tcctggtggt ctcaagcggc acgtgattga agacgagcta
     2041 gtgttattcc tgactgtgaa accaaagcgg agtgactgca actaatgaaa attaaatttt
     2101 taaggaagga tagagactcc caactgttcg tgcaccatga gagcaaggat gcccatttgt
     2161 gccgcacgtc cctggttgag ttcgatagca cgcttctgaa gcttcttgtc ctcactgaaa
     2221 gtgtcccagc caaagtcgag gtagttgttg cggccgcatg ggcccagatc tccgaggcct
     2281 cggacccgtc gggccgcgtc ggaccggcgg tgttggtcgg cgtcggtcag tcctgctcct
     2341 cggccacgaa gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg
     2401 gctgctcgcc gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga
     2461 cctccgacca ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt
     2521 tgtccggcac cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca
     2581 caccggcgaa gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact
     2641 cgaccgctcc ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca
     2701 tgggtaccct cagctagaat attatcgata ccgtcgactt ggttaatttt tcgatttcga
     2761 agatatgtga agatccaggt ggtgtagatc acgaaagcgt tccgccgcgt gccaatccat
     2821 ccatcggcag ggatcgtcgt tcaacctgcc tacacgtgct tcgcacgtag aaattctacc
     2881 agtacaacga aagctgctgt ccgacgttac gaacatggtc tagtcagaca gttgaaagac
     2941 ccggagtttt gagatcttcg gtcgatcata aaaaaccttg ctagcacaga aatattgctg
     3001 agttacgttc acaccaaaaa cagagcactg ttccaagtcc gcaagttact tttacatgta
     3061 aaagcgttgt ctatgtttat gaaatcagca ccgcatgtcg aaaatgctat tttgagaatc
     3121 gccgatgcta atcattggcc tctaaaaaaa cggaagtatc aagaattcca gacaaatcag
     3181 gatttacaga taacataaat cgcgaaatgg aattttgcaa ccaacgttaa caggtatgtc
     3241 tccttaccaa acgctagtcc agtccctccc agctgatgtc tttcaaccaa caagtatcta
     3301 cgactgcagc actaatgcaa atgataatat aaaaaagaag acaatgtttt tcccagtgat
     3361 tcaactcgaa ttgcactaat cctgttaggt accagctttt gttcccttta gtgagggtta
     3421 atttcgagct tggcgtaatc atggtaagct gagacctgag acgagacgag acagcctgag
     3481 aatggat
//
