LOCUS       sequence-84641-e        3243 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 3243)
  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 3243)
  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 D-E mOrange + link c2
FEATURES             Location/Qualifiers
     source          1..3243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(147..152,153..169,170..175)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             364..897
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     984..1002
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1040..1062)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1162..1181
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1375..1397
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1389..1406
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1390..1406
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1535..2242
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGF
                     QTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2285..2301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2285..2301)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2298..2320)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2309..2325
                     /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(2333..2339,2340..2357,2358..2363))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2378..2399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2516..2533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2687..3243,1..32))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2767..2786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
       61 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgagg cgcacaccgt
      121 ggaaacggat gaaggcacga acccagttga cataagcctg ttcggttcgt aaactgtaat
      181 gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
      241 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag tctatgcctc
      301 gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca
      361 acgatgttac gcagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
      421 ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca agtcaaatcc
      481 atgcgggctg ctcttgatct tttcggtcgt gagttcggcg acgtagccac ctactcccaa
      541 catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt catcgcgctt
      601 gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct gcccaggttt
      661 gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggcga gcaccggagg
      721 cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc gcttggtgct
      781 tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct ctatacaaag
      841 ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc cacctaacaa
      901 ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt tcggtaaatt gtcacaacgc
      961 cgctcatgac attaacctat aaaaataggc gtatcacgag gcccttttgt ctcgcgcgtt
     1021 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagaaggtc acagcttgtc
     1081 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     1141 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     1201 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     1261 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     1321 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     1381 cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacctcgcga atgcatctag
     1441 atatcggatc ccgggcccgt cgactgcaga ggtacgggtc tcaaagggag gtggaggagg
     1501 ttctggaggc ggtggaagtg gtggcggagg tagcgtgagc aagggcgagg agaataacat
     1561 ggccatcatc aaggagttca tgcgcttcaa ggtgcgcatg gagggctctg tgaacggcca
     1621 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggctttc agaccgctaa
     1681 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1741 cacctacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttcaagct
     1801 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1861 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1921 cggcaccaac ttcccctccg acggccccgt aatgcagaag aaaaccatgg gctgggaggc
     1981 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct
     2041 gaagctgaag gacggcggcc actacacctc cgaggtcaag accacctaca aggccaagaa
     2101 gcccgtgcag ctgcccggcg cctacatcgt cggcatcaag ttggacatca cctcccacaa
     2161 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     2221 catggacgag ctgtacaagt aaaatctgag acctctgcct gcatgcaagc ttggcgtaat
     2281 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2341 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2401 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2461 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3121 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3241 agc
//
