LOCUS       sequence-75080-e        2959 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 2959)
  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 2959)
  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: pICSL80007
FEATURES             Location/Qualifiers
     source          1..2959
                     /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             2249..2959
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2394..2412)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
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 ggtgagcaag ggcgaggagg ataacatggc
     2281 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     2341 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2401 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     2461 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2521 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     2581 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2641 caccaacttc ccctccgacg gcccagtaat gcagaagaaa accatgggct gggaggcctc
     2701 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     2761 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2821 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     2881 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     2941 ggacgagctg tacaagtga
//
