LOCUS       sequence-497554-        2970 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 2970)
  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 2970)
  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: pICSL80014
FEATURES             Location/Qualifiers
     source          1..2970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      197..785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     686..705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     939..956
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(1034..1036,1037..1039,1040..1753)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1079..1100)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1340..1359)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1687..1708
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1793..1811)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1879..1983
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2205..2970,1..26)
                     /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 agatcaccaa ggtagtcggc aaataactgt cagaccaagt ttactcatat atactttaga
       61 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
      121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      781 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      841 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      901 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      961 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatcac
     1021 tctgtggtct caaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1081 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1141 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1201 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
     1261 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1321 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1381 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1441 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1501 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1561 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1621 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     1681 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1741 gctgtacaag tgagctttga gaccacgaag tggctcttca gtggacgaaa gggcctcgtg
     1801 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     1861 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1921 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1981 agtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     2041 gtggcggttt tcatggcttg ttatgactgt ttttttttgg ggtacagtct atgcctcggg
     2101 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     2161 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg gaagcggtga
     2221 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     2281 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     2341 gcgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     2401 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     2461 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     2521 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     2581 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     2641 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     2701 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     2761 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     2821 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     2881 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     2941 tggaagaatt tgtccattac gtaaaaggcg
//
