LOCUS       sequence-84650-e        2588 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 2588)
  AUTHORS   Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske 
            M
  TITLE     A modular plasmid assembly kit for multigene expression, gene 
            silencing and silencing rescue in plants.
  JOURNAL   PLoS One. 2014 Feb 13;9(2):e88218. doi: 
            10.1371/journal.pone.0088218. eCollection 2014.
  PUBMED    24551083
REFERENCE   2  (bases 1 to 2588)
  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"; journalName: "PLoS One.";
            date: "2014-02-13"; volume: "9(2):e88218. doi"; pages: " 
            10.1371/journal.pone.0088218. eCollection 2014"
COMMENT     Sequence Label: LI D-E 11GFP
FEATURES             Location/Qualifiers
     source          1..2588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     466..484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(522..544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     644..663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     857..879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     871..888
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     872..888
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             925..948
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             976..1023
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     primer_bind     complement(1112..1128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1125..1147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1136..1152
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1160..1166,1167..1184,1185..1190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1205..1226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1343..1360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1514..2102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1594..1613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2217..2222,2223..2239,2240..2245)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2434..2588,1..379)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cgacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
       61 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
      121 ggcttacgtt ctgcccaggt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
      181 agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
      241 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
      301 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
      361 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
      421 gttcggtaaa ttgtcacaac gccgctcatg acattaacct ataaaaatag gcgtatcacg
      481 aggccctttt gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
      541 ccggagaagg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      601 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      661 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      721 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      781 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      841 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
      901 ggtacctcga tggtctcaaa gggagactac aaggacgacg atgacaaggg cgacggcggc
      961 agcggcggcg gcagcaggga ccacatggtt ctccacgaat acgtcaacgc cgccggaatc
     1021 acataaaatc agagacctac gaatgcatct agatatcgga tcccgggccc gtcgactgca
     1081 gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1141 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1201 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1321 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1381 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2101 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2161 aagggatttt ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga acccagttga
     2221 cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt
     2281 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2341 atgactgttt ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     2401 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg
     2461 cagcagggca gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat cattcgcaca
     2521 tgtaggctcg gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt
     2581 gagttcgg
//
