LOCUS       sequence-405601-        2773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a sfGFP fluorescent protein with CD 
            overhangs used to build a level 1 construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2773)
  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 2773)
  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_CD-sfGFP
FEATURES             Location/Qualifiers
     source          1..2773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1072..1785
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     rep_origin      complement(1930..2518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2010..2029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2609..2773,1..627))
                     /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 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
       61 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
      121 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
      181 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
      241 aatgtcgatc gtggctggct cgaagatacc agcaagaatg tcattgcgct gccattctcc
      301 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
      361 ggtgacttct acagcgcgga gaatctcact ctctccaggg gaagccgaag tttccaaaag
      421 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
      481 atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
      541 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
      601 tacttcggcg atcaccgctt ccctcatgcg aaacgatcct catcctgtct cttgatcaga
      661 tattgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca
      721 gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc ttgctaagct
      781 tgcatgcctg caagtcgact ctagaggagc ggtgatcaca ggcagcaacg ctctgtcatc
      841 gttacaatca acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt
      901 tgccgttctt ccgaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct
      961 cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
     1021 ctcattactc gcatccattc tcaggctgtc tcgtctcgtc tcaggtctca aatgcgtaaa
     1081 ggcgaggagc tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac
     1141 ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg
     1201 ctgaagttca tctgtactac tggtaaactg ccggtacctt ggccgactct ggtaacgacg
     1261 ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc
     1321 ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac
     1381 ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt
     1441 gagctgaaag gcattgactt taaagaagac ggcaatatcc tgggccataa gctggaatac
     1501 aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
     1561 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag
     1621 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
     1681 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
     1741 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatcagg tcgagaccac
     1801 caggatacat agattaccac aactccgagc ccttccaccc acagaatcag gggataacgc
     1861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2341 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2581 gattttggtc atgcattcta ggtgattatt atttgccgac taccttggtg atctcgcctt
     2641 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
     2701 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
     2761 ccattgccca gtc
//
