LOCUS       sequence-497541-        2529 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 2529)
  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 2529)
  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: pICH42211
FEATURES             Location/Qualifiers
     source          1..2529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     123..142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     376..393
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        550..729
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(789..807)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        875..979
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1201..1992
                     /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"
     rep_origin      join(2163..2529,1..222)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
       61 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      301 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      361 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      421 ccgcctctcc ccgcgcgttg gccgattcat taatcactct gtggtctcag gagagcggag
      481 aattaaggga gtcacgttat gacccccgcc gatgacgcgg gacaagccgt tttacgtttg
      541 gaactgacag aaccgcaacg ttgaaggagc cactcagccg cgggtttctg gagtttaatg
      601 agctaagcac atacgtcaga aaccattatt gcgcgttcaa aagtcgccta aggtcactat
      661 cagctagcaa atatttcttg tcaaaaatgc tccactgacg ttccataaat tcccctcggt
      721 atccaattag agtctcatat tcactctcat acttgagacc acgaagtggc tcttcagtgg
      781 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
      841 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      901 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      961 atattgaaaa aggaagagta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     1021 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttttggggta
     1081 cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg
     1141 ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatc
     1201 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
     1261 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
     1321 ggcctgaagc cacacagcga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
     1381 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
     1441 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
     1501 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
     1561 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
     1621 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
     1681 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
     1741 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
     1801 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
     1861 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
     1921 tcgcgcgcag atcagttgga agaatttgtc cattacgtaa aaggcgagat caccaaggta
     1981 gtcggcaaat aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2041 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2101 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2161 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2221 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2281 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2341 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2401 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2461 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2521 acctacacc
//
