LOCUS       sequence-84646-e        3247 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 3247)
  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 3247)
  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 mCherry
FEATURES             Location/Qualifiers
     source          1..3247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     587..605
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(643..665)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     765..784
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     978..1000
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     992..1009
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     993..1009
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1095..1802
                     /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(1240..1258)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1892..1908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1892..1908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1905..1927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1916..1932
                     /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(1940..1946,1947..1964,1965..1970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1985..2006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2123..2140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2294..2882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2374..2393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2997..3002,3003..3019,3020..3025)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3214..3247,1..500)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gcagtcgccc taaaacaaag ttaggtggct caagtatggg catcattcgc acatgtaggc
       61 tcggccctga ccaagtcaaa tccatgcggg ctgctcttga tcttttcggt cgtgagttcg
      121 gcgacgtagc cacctactcc caacatcagc cggactccga ttacctcggg aacttgctcc
      181 gtagtaagac attcatcgcg cttgctgcct tcgaccaaga agcggttgtt ggcgctctcg
      241 cggcttacgt tctgcccagg tttgagcagc cgcgtagtga gatctatatc tatgatctcg
      301 cagtctccgg cgagcaccgg aggcagggca ttgccaccgc gctcatcaat ctcctcaagc
      361 atgaggccaa cgcgcttggt gcttatgtga tctacgtgca agcagattac ggtgacgatc
      421 ccgcagtggc tctctataca aagttgggca tacgggaaga agtgatgcac tttgatatcg
      481 acccaagtac cgccacctaa caattcgttc aagccgagat cggcttcccg gccgcggagt
      541 tgttcggtaa attgtcacaa cgccgctcat gacattaacc tataaaaata ggcgtatcac
      601 gaggcccttt tgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
      661 cccggagaag gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      721 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      781 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      841 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      901 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      961 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
     1021 cggtacctcg cgaatgcatc tagatatcgg atcccgggcc cgtcgactgc agaggtacgg
     1081 gtctcatctg aacaatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     1141 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1201 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1261 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1321 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1381 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1441 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     1501 ccgacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc gagcggatgt
     1561 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     1621 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     1681 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     1741 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     1801 agggtggagg aggttctgga ggcggtggaa gtggtggcgg aggtagcggc acctgagacc
     1861 tctgcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1921 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1981 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2281 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2341 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2401 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2461 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2521 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2581 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2641 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2701 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2761 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2821 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2881 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2941 aagggatttt ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga acccagttga
     3001 cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca cgcaactggt
     3061 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     3121 atgactgttt ttttgtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     3181 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagcaa cgatgttacg
     3241 cagcagg
//
