LOCUS       sequence-405621-        2783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a Venus 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 2783)
  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 2783)
  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-Venus
FEATURES             Location/Qualifiers
     source          1..2783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      433..1021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     922..941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1167..1880,1881..1883,1884..1886))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1212..1233)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1561..1580
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1820..1841
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             join(2334..2783,1..342)
                     /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 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
       61 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
      121 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
      181 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
      241 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
      301 cactacgtga aaggcgagat caccaaggta gtcggcaaat aataatcacc tagaatgcat
      361 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
      421 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
      481 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      541 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      601 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      661 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      721 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      781 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      841 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      901 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      961 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1021 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1081 gttctttcct gcgttatccc ctgattctgt gggtggaagg gctcggagtt gtggtaatct
     1141 atgtatcctg gtggtctcta agcttatcac ttgtacagct cgtccatgcc gagagtgatc
     1201 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
     1261 ctcagggcgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
     1321 atgggggtgt tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg
     1381 cggatcttga agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg
     1441 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     1501 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     1561 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     1621 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     1681 aggccgtagc ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     1741 cagatcagct tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg
     1801 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     1861 aacagctcct cgcccttgct caccatacct tgagacctga gacgagacga gacagcctga
     1921 gaatggatgc gagtaatgag ctcggcacaa aatcaccact cgatacaggc agcccatcag
     1981 tccgggacgg cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga
     2041 tgctattcgg aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag
     2101 ggtagcatgt tgattgtaac gatgacagag cgttgctgcc tgtgatcacc gctcctctag
     2161 agtcgacttg caggcatgca agcttagcaa gcgaaccgga attgccagct ggggcgccct
     2221 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     2281 gatggcgcag gggatcaata tctgatcaag agacaggatg aggatcgttt cgcatgaggg
     2341 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
     2401 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
     2461 agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
     2521 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agtgagattc
     2581 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
     2641 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgct ggtatcttcg
     2701 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
     2761 ttgccttggt aggtccagcg gcg
//
