LOCUS       sequence-84636-e        3205 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 3205)
  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 3205)
  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 C-D T-Saphire c1
FEATURES             Location/Qualifiers
     source          1..3205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..556
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     643..661
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(699..721)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     821..840
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1034..1056
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1048..1065
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1049..1065
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1147..1149,1150..1152,1153..1863)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1192..1213)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1453..1472)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1800..1821
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1906..1922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1906..1922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1919..1941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1930..1946
                     /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(1954..1960,1961..1978,1979..1984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1999..2020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2137..2154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2308..2896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2388..2407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3011..3016,3017..3033,3034..3039)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ttgatgttat ggagcagcaa cgatgttacg cagcagcaac gatgttacgc agcagggcag
       61 tcgccctaaa acaaagttag gtggctcaag tatgggcatc attcgcacat gtaggctcgg
      121 ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt ttcggtcgtg agttcggcga
      181 cgtagccacc tactcccaac atcagccgga ctccgattac ctcgggaact tgctccgtag
      241 taagacattc atcgcgcttg ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc
      301 ttacgttctg cccaggtttg agcagccgcg tagtgagatc tatatctatg atctcgcagt
      361 ctccggcgag caccggaggc agggcattgc caccgcgctc atcaatctcc tcaagcatga
      421 ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg acgatcccgc
      481 agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg atatcgaccc
      541 aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttcccggccg cggagttgtt
      601 cggtaaattg tcacaacgcc gctcatgaca ttaacctata aaaataggcg tatcacgagg
      661 cccttttgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      721 gagaaggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      781 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      841 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      901 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      961 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     1021 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt
     1081 acctcgcgaa tgcatctaga tatcggatcc cgggcccgtc gactgcagag gtacgggtct
     1141 cacaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1201 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1261 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1321 cccaccctcg tgaccacctt ctcctacggc gtgatggtgt tcgcccgcta ccccgaccac
     1381 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1441 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1501 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1561 gggcacaagc tggagtacaa cttcaacagc cacaacgtct atatcatggc cgacaagcag
     1621 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     1681 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1741 aaccactacc tgagcatcca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1801 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1861 aagaaggtga gacctctgcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     1921 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1981 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2041 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2101 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2161 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2221 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2281 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2341 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2641 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2701 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2761 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2821 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2941 cgaaaactca cgttaaggga ttttggtcat gaggcgcaca ccgtggaaac ggatgaaggc
     3001 acgaacccag ttgacataag cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg
     3061 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     3121 tttcatggct tgttatgact gtttttttgt acagtctatg cctcgggcat ccaagcagca
     3181 agcgcgttac gccgtgggtc gatgt
//
