LOCUS       sequence-84651-e        2589 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 2589)
  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 2589)
  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 11GFP
FEATURES             Location/Qualifiers
     source          1..2589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(344..349,350..366,367..372))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      487..1075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     976..995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1229..1246
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1363..1384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1399..1404,1405..1422,1423..1429)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1437..1453
                     /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)."
     primer_bind     1442..1464
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1461..1477
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1461..1477
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1566..1589
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1617..1664
                     /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(1702..1719)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1702..1718)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1711..1733)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1927..1946)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2046..2068
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2106..2124)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(2211..2589,1..155))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
       61 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
      121 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
      181 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
      241 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
      301 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
      361 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgcc tcatgaccaa
      421 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      481 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      541 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      601 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      661 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      721 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      781 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      841 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      901 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      961 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1021 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1081 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1141 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1201 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1261 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1321 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     1381 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     1441 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg
     1501 caggcctctg cagtcgacgg gcccgggatc cgatatctag atgcattcga tggtctcaca
     1561 ccatggacta caaggacgac gatgacaagg gcgacggcgg cagcggcggc ggcagcaggg
     1621 accacatggt tctccacgaa tacgtcaacg ccgccggaat cacataaaag gagagaccta
     1681 cgaggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1741 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1801 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     1861 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     1921 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1981 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2041 cttctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagaca
     2101 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgagcggcg ttgtgacaat
     2161 ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg
     2221 gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga
     2281 gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg
     2341 ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg
     2401 ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa cctgggcaga
     2461 acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat
     2521 gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg
     2581 gctacgtcg
//
