LOCUS       sequence-497577-        2972 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2972)
  AUTHORS   Engler C, Youles M, Gruetzner R, Ehnert TM, Werner S, Jones JD, 
            Patron NJ, Marillonnet S
  TITLE     A golden gate modular cloning toolbox for plants.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):839-43. doi: 10.1021/sb4001504. 
            Epub 2014 Feb 20.
  PUBMED    24933124
REFERENCE   2  (bases 1 to 2972)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/sb4001504";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "839-43"
COMMENT     Sequence Label: pICSL80012
FEATURES             Location/Qualifiers
     source          1..2972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(734..838)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     906..924
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(964..1677,1678..1680))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1009..1030)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1358..1377
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1617..1638
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1763..1780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1934..2522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2014..2033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2693..2972,1..512))
                     /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 gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc
       61 tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc
      121 agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat
      181 tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca
      241 acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc
      301 aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc
      361 agcaaatcaa tatcgctgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt
      421 acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga
      481 gtcgatactt cggcgatcac cgcttccctc atgatgttta actttgtttt agggcgactg
      541 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
      601 gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaaaacag tcataacaag
      661 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
      721 tgcgtgagcg catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
      781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
      841 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
      901 ataaaaatag gcgtatcacg aggccctttc gtccactgaa gagccacttc gtggtctcaa
      961 agctcacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     1021 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact ggtagctcag
     1081 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     1141 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     1201 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     1261 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     1321 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     1381 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     1441 ctgcttcatg tggtcggggt agcgggcgaa gcactgcagg ccgtagccga aggtggtcac
     1501 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     1561 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     1621 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     1681 taaccttgag accacagagt gattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1741 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1801 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1861 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1921 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1981 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2041 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2101 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2161 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2221 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2281 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2341 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2401 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2461 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2521 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2581 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2641 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttatttgc
     2701 cgactacctt ggtgatctcg ccttttacgt aatggacaaa ttcttccaac tgatctgcgc
     2761 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
     2821 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
     2881 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
     2941 cgccagccca gtcgggcggc gagttccata gc
//
