LOCUS       sequence-405600-        2776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a mTurquoise2 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 2776)
  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 2776)
  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-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..2776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      819..1407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1308..1327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1552..2262,2263..2265,2266..2268))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1594..1615)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1943..1962
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2202..2223
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             join(2713..2776,1..728)
                     /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 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
       61 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
      121 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagtgaga
      181 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
      241 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gctggtatct
      301 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
      361 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
      421 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
      481 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
      541 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
      601 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
      661 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
      721 gcaaataata atcacctaga atgcatgacc aaaatccctt aacgtgagtt ttcgttccac
      781 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      961 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1381 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1441 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtgggt
     1501 ggaagggctc ggagttgtgg taatctatgt atcctggtgg tctctacctg acttgtacag
     1561 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     1621 gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg
     1681 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     1741 gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc ttgatgccgt tcttctgctt
     1801 gtcggcggtg atatagacgt tgtcgctaaa gtagttgtac tccagcttgt gccccaggat
     1861 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     1921 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     1981 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     2041 ggggtagcgg gcgaagcact gcacgcccca ggacagggtg gtcacgaggg tgggccaggg
     2101 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     2161 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     2221 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccattt gagacctgag
     2281 acgagacgag acagcctgag aatggatgcg agtaatgagc tcggcacaaa atcaccactc
     2341 gatacaggca gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag
     2401 actttgctca tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa
     2461 cacggatgat ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct
     2521 gtgatcaccg ctcctctaga gtcgacttgc aggcatgcaa gcttagcaag cgaaccggaa
     2581 ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct
     2641 ttcttgccgc caaggatctg atggcgcagg ggatcaatat ctgatcaaga gacaggatga
     2701 ggatcgtttc gcatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
     2761 gttggcgtca tcgagc
//
