LOCUS       sequence-84591-e        4252 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 4252)
  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 4252)
  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: LIIc F 4-5
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     273..295
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     287..304
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     288..304
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     315..334
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        315..333
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    474..598
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(635..640,641..658,659..665)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             719..1378
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(785..804)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1720..2025
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1904..1923
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2066..2190)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2390..2407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2500..3088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2580..2599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3259..4050,4051..4119))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3882..3901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4120..4224)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
       61 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
      121 ggttgagtgg ccgctacagg gcgctcccat tcgccattca ggctgcgcaa ctgttgggaa
      181 gggcgtttcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      241 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      301 cagtgagcgc gacgtaatac gactcactat agggcgaatt ggcggaaggc cgtcaaggcc
      361 acgtgtcttg tccaggcgcg ccatgagctc taaagcttcg ctcgagctgc ggccgcacgg
      421 atcctcgaat tcccaagctt atctcgaccc gaagacaatg tcgcggtgag accacaagtt
      481 tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
      541 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggc
      601 ggcccctcga ggggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta
      661 taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat
      721 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
      781 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
      841 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
      901 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
      961 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     1021 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     1081 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     1141 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     1201 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     1261 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     1321 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatc
     1381 tagaggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg
     1441 cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg
     1501 aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg
     1561 atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg
     1621 ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa
     1681 tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc
     1741 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg
     1801 cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtcccc
     1861 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat
     1921 atggccagtg tgccggtttc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa
     1981 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt
     2041 atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt tacagtatta
     2101 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     2161 ttctcgttca gctttcttgt acaaagtggt tggggtctca tgtctgagtt gtcttcgggg
     2221 tagcggccgc ttggatccca gaattctagg tacctcttaa ttaatggagc acaagactgg
     2281 cctcatgggc cttccgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2341 attaacatgg tcatagctgt ttccttgcgt attgggcgct ctccgcttcc tcgctcactg
     2401 actcgctgcg ctcggtcgtt cgggtaaagc ctggggtgcc taatgagcaa aaggccagca
     2461 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2521 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2581 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2641 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2701 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2761 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2821 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2881 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2941 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3001 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3061 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3121 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3181 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3241 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3301 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3361 gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc
     3421 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3481 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3541 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3601 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3661 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3721 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3781 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3841 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3901 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3961 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4021 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4081 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4141 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4201 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aa
//
