LOCUS       sequence-84647-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 mOrange
FEATURES             Location/Qualifiers
     source          1..3247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     547..565
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(603..625)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     725..744
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     938..960
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     952..969
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     953..969
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1055..1762
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1852..1868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1852..1868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1865..1887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1876..1892
                     /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(1900..1906,1907..1924,1925..1930))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1945..1966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2083..2100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2254..2842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2334..2353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2957..2962,2963..2979,2980..2985)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3174..3247,1..460)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg ctgctcttga
       61 tcttttcggt cgtgagttcg gcgacgtagc cacctactcc caacatcagc cggactccga
      121 ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct tcgaccaaga
      181 agcggttgtt ggcgctctcg cggcttacgt tctgcccagg tttgagcagc cgcgtagtga
      241 gatctatatc tatgatctcg cagtctccgg cgagcaccgg aggcagggca ttgccaccgc
      301 gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga tctacgtgca
      361 agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca tacgggaaga
      421 agtgatgcac tttgatatcg acccaagtac cgccacctaa caattcgttc aagccgagat
      481 cggcttcccg gccgcggagt tgttcggtaa attgtcacaa cgccgctcat gacattaacc
      541 tataaaaata ggcgtatcac gaggcccttt tgtctcgcgc gtttcggtga tgacggtgaa
      601 aacctctgac acatgcagct cccggagaag gtcacagctt gtctgtaagc ggatgccggg
      661 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
      721 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
      781 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
      841 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      901 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      961 acggccagtg aattcgagct cggtacctcg cgaatgcatc tagatatcgg atcccgggcc
     1021 cgtcgactgc agaggtacgg gtctcatctg aacaatggtg agcaagggcg aggagaataa
     1081 catggccatc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ctgtgaacgg
     1141 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggct ttcagaccgc
     1201 taagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     1261 gttcacctac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttcaa
     1321 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     1381 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     1441 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaaaacca tgggctggga
     1501 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagatgag
     1561 gctgaagctg aaggacggcg gccactacac ctccgaggtc aagaccacct acaaggccaa
     1621 gaagcccgtg cagctgcccg gcgcctacat cgtcggcatc aagttggaca tcacctccca
     1681 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     1741 cggcatggac gagctgtaca agggtggagg aggttctgga ggcggtggaa gtggtggcgg
     1801 aggtagcggc acctgagacc tctgcctgca tgcaagcttg gcgtaatcat ggtcatagct
     1861 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1921 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1981 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2041 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2101 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2161 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2221 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2281 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2341 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2401 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2461 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2521 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2581 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2641 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2701 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2761 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2821 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2881 gtggaacgaa aactcacgtt aagggatttt ggtcatgagg cgcacaccgt ggaaacggat
     2941 gaaggcacga acccagttga cataagcctg ttcggttcgt aaactgtaat gcaagtagcg
     3001 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     3061 ggcggttttc atggcttgtt atgactgttt ttttgtacag tctatgcctc gggcatccaa
     3121 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     3181 gcagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggtggctcaa
     3241 gtatggg
//
