LOCUS       sequence-405620-        2774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a sfGFP 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 2774)
  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 2774)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..2774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(141..160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(740..1531)
                     /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"
     CDS             1979..2689
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
ORIGIN
        1 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
       61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      481 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      721 catgcattct aggtgattat tatttgccga ctaccttggt gatctcgcct ttcacgtagt
      781 ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat
      841 aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc
      901 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg
      961 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg
     1021 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct
     1081 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca
     1141 gatcaatgtc gatcgtggct ggctcgaaga taccagcaag aatgtcattg cgctgccatt
     1201 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa
     1261 caatggtgac ttctacagcg cggagaatct cactctctcc aggggaagcc gaagtttcca
     1321 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca
     1381 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta
     1441 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag
     1501 tcgatacttc ggcgatcacc gcttccctca tgcgaaacga tcctcatcct gtctcttgat
     1561 cagatattga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     1621 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgcta
     1681 agcttgcatg cctgcaagtc gactctagag gagcggtgat cacaggcagc aacgctctgt
     1741 catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct
     1801 tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg
     1861 ctctcccgct gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc
     1921 cgagctcatt actcgcatcc attctcaggc tgtctcgtct cgtctcaggt ctcaaggtcg
     1981 taaaggcgag gagctgttca ctggtgtcgt ccctattctg gtggaactgg atggtgatgt
     2041 caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta atggtaaact
     2101 gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga ctctggtaac
     2161 gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga agcagcatga
     2221 cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt cctttaagga
     2281 tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg
     2341 cattgagctg aaaggcattg actttaaaga agacggcaat atcctgggcc ataagctgga
     2401 atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa atggcattaa
     2461 agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg ctgatcacta
     2521 ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc actatctgag
     2581 cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg ttctgctgga
     2641 gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat aagcttagag
     2701 accaccagga tacatagatt accacaactc cgagcccttc cacccacaga atcaggggat
     2761 aacgcaggaa agaa
//
