LOCUS       sequence-99963-e        2544 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 2544)
  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 2544)
  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 B-C N-NLS KOZ c2
FEATURES             Location/Qualifiers
     source          1..2544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     350..368
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(406..428)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     528..547
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     741..763
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     755..772
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     756..772
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(833..853)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(952..968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(952..968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(965..987)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    976..992
                     /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(1000..1006,1007..1024,1025..1030))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1045..1066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1183..1200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1354..1942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1434..1453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2057..2062,2063..2079,2080..2085)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2274..2544,1..263)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 tcgcggctta cgttctgccc aggtttgagc agccgcgtag tgagatctat atctatgatc
       61 tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc aatctcctca
      121 agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat tacggtgacg
      181 atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg cactttgata
      241 tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc ccggccgcgg
      301 agttgttcgg taaattgtca caacgccgct catgacatta acctataaaa ataggcgtat
      361 cacgaggccc ttttgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
      421 gctcccggag aaggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
      481 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
      541 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
      601 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
      661 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
      721 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcga
      781 gctcggtacc tcgtaggtct caggtggcgg ccgcactcga gtcgactcct ccaacctttc
      841 tcttcttctt aggctgcagc attgttcaga tgagaccatc gaatgcatct agatatcgga
      901 tcccgggccc gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct
      961 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1021 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1081 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1141 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1201 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1261 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1321 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1381 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1441 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1501 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1561 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1621 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1681 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1741 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1801 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1861 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1981 gtggaacgaa aactcacgtt aagggatttt ggtcatgagg cgcacaccgt ggaaacggat
     2041 gaaggcacga acccagttga cataagcctg ttcggttcgt aaactgtaat gcaagtagcg
     2101 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     2161 ggcggttttc atggcttgtt atgactgttt ttttgtacag tctatgcctc gggcatccaa
     2221 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     2281 gcagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggtggctcaa
     2341 gtatgggcat cattcgcaca tgtaggctcg gccctgacca agtcaaatcc atgcgggctg
     2401 ctcttgatct tttcggtcgt gagttcggcg acgtagccac ctactcccaa catcagccgg
     2461 actccgatta cctcgggaac ttgctccgta gtaagacatt catcgcgctt gctgccttcg
     2521 accaagaagc ggttgttggc gctc
//
