LOCUS       sequence-405610-        2781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a Venus 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 2781)
  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 2781)
  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-Venus
FEATURES             Location/Qualifiers
     source          1..2781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(411..413,414..416,417..1127)
                     /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(456..477)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(717..736)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1064..1085
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      complement(1272..1860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1352..1371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1951..2742)
                     /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 tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt
       61 cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgcta agcttgcatg
      121 cctgcaagtc gactctagag gagcggtgat cacaggcagc aacgctctgt catcgttaca
      181 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
      241 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
      301 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctcatt
      361 actcgcatcc attctcaggc tgtctcgtct cgtctcgtct caggtctcaa atggtgagca
      421 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      481 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      541 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      601 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
      661 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      721 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      781 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      841 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
      901 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
      961 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1021 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1081 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtca ggtagagacc
     1141 accaggatac atagattacc acaactccga gcccttccac ccacagaatc aggggataac
     1201 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1261 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1321 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1381 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1441 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1501 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1561 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1621 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1681 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1741 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1801 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1861 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1921 gggattttgg tcatgcattc taggtgatta ttatttgccg actaccttgg tgatctcgcc
     1981 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc
     2041 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
     2101 ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta
     2161 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
     2221 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
     2281 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
     2341 caagatagcc agatcaatgt cgatcgtggc tggctcgaag ataccagcaa gaatgtcatt
     2401 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     2461 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcactctctc caggggaagc
     2521 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     2581 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     2641 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     2701 tgatagttga gtcgatactt cggcgatcac cgcttccctc atgcgaaacg atcctcatcc
     2761 tgtctcttga tcagatattg a
//
