LOCUS       sequence-497570-        2964 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 2964)
  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 2964)
  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: pICSL80005
FEATURES             Location/Qualifiers
     source          1..2964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1148..1165)
                     /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"
     primer_bind     complement(1213..1231)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1299..1403
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1625..2416
                     /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(2587..2964,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatcactctg tggtctcaaa tgagaggatc tggatctgag
      481 tctgatgagt ctggacttcc tgctatggaa atcgagtgta gaatcactgg aacccttaac
      541 ggtgttgagt tcgagcttgt tggaggtggt gagggaactc ctgagcaggg aagaatgact
      601 aacaagatga agtctaccaa gggtgctctt accttctctc cataccttct ttctcacgtt
      661 atgggatacg gattctacca cttcggaact tacccatctg gatacgagaa ccctttcctt
      721 catgctatca acaacggtgg atacaccaac actaggatcg agaagtacga ggatggtggt
      781 gttcttcacg ttagcttctc ttacagatac gaggctggaa gagtgatcgg agatttcaag
      841 gttatgggaa ctggattccc tgaggattct gttatcttca ccgacaagat catcaggtct
      901 aacgctactg ttgagcatct tcatcctatg ggagataacg atctcgatgg atctttcacc
      961 agaaccttct cacttagaga tggtggttac tactcttctg tggtggattc tcacatgcac
     1021 ttcaagtctg ctatccaccc ttctatcctt caaaacggtg gacctatgtt cgctttcaga
     1081 agagttgagg aagatcactc taacaccgag cttggaatcg ttgagtacca acatgctttc
     1141 aagacccctg atgctgatgc tggtgaggaa taggctttga gaccacgaag tggctcttca
     1201 gtggacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1261 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1321 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1381 aataatattg aaaaaggaag agtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     1441 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttttgg
     1501 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     1561 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     1621 catcatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     1681 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     1741 tggcggcctg aagccacaca gcgatattga tttgctggtt acggtgaccg taaggcttga
     1801 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     1861 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     1921 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     1981 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     2041 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     2101 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     2161 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     2221 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     2281 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     2341 ggcctcgcgc gcagatcagt tggaagaatt tgtccattac gtaaaaggcg agatcaccaa
     2401 ggtagtcggc aaataactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2461 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2521 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2581 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2641 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2701 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2761 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2821 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2881 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2941 aacgacctac accgaactga gata
//
