LOCUS       sequence-75079-e        2962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2962)
  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 2962)
  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"; doi: "10.1021/sb4001504";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "839-43"
COMMENT     Sequence Label: pICSL80005
FEATURES             Location/Qualifiers
     source          1..2962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..58)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        126..230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             450..1241
                     /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      1412..2000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1901..1920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2154..2171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(2937..2954)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
ORIGIN
        1 gctttgagac cacgaagtgg ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt
       61 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgcgctcac
      241 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
      301 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
      361 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
      421 gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga
      481 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
      541 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc
      601 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
      661 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
      721 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
      781 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
      841 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
      901 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
      961 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     1021 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     1081 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     1141 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     1201 attacgtaaa aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact
     1261 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1321 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1381 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1441 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1501 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1561 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1621 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1681 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1741 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1801 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1861 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1921 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1981 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2041 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2101 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2161 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2221 cgattcatta atcactctgt ggtctcaaat gagaggatct ggatctgagt ctgatgagtc
     2281 tggacttcct gctatggaaa tcgagtgtag aatcactgga acccttaacg gtgttgagtt
     2341 cgagcttgtt ggaggtggtg agggaactcc tgagcaggga agaatgacta acaagatgaa
     2401 gtctaccaag ggtgctctta ccttctctcc ataccttctt tctcacgtta tgggatacgg
     2461 attctaccac ttcggaactt acccatctgg atacgagaac cctttccttc atgctatcaa
     2521 caacggtgga tacaccaaca ctaggatcga gaagtacgag gatggtggtg ttcttcacgt
     2581 tagcttctct tacagatacg aggctggaag agtgatcgga gatttcaagg ttatgggaac
     2641 tggattccct gaggattctg ttatcttcac cgacaagatc atcaggtcta acgctactgt
     2701 tgagcatctt catcctatgg gagataacga tctcgatgga tctttcacca gaaccttctc
     2761 acttagagat ggtggttact actcttctgt ggtggattct cacatgcact tcaagtctgc
     2821 tatccaccct tctatccttc aaaacggtgg acctatgttc gctttcagaa gagttgagga
     2881 agatcactct aacaccgagc ttggaatcgt tgagtaccaa catgctttca agacccctga
     2941 tgctgatgct ggtgaggaat ag
//
