LOCUS       sequence-84589-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 3-4
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     253..275
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     267..284
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     268..284
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     295..314
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        295..313
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    454..578
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(615..620,621..638,639..645)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             699..1358
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(765..784)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1700..2005
                     /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     1884..1903
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2046..2170)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2370..2387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2480..3068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2560..2579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3239..4030,4031..4099))
                     /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     3862..3881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4100..4204)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
       61 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgg ccgctacagg
      121 gcgctcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgtttcg gtgcgggcct
      181 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      241 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gacgtaatac
      301 gactcactat agggcgaatt ggcggaaggc cgtcaaggcc acgtgtcttg tccaggcgcg
      361 ccatgagctc taaagcttcg ctcgagctgc ggccgcacgg atcctcgaat tcccaagctt
      421 atctcgaccc gaagacaaat tggcggtgag accacaagtt tgtacaaaaa agctgaacga
      481 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
      541 acataatact gtaaaacaca acatatccag tcactatggc ggcccctcga ggggccgcat
      601 taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt
      661 aggatccgtc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
      721 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
      781 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
      841 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
      901 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
      961 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1021 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     1081 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     1141 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     1201 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     1261 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     1321 tacaacagta ctgcgatgag tggcagggcg gggcgtaatc tagaggatcc ggcttactaa
     1381 aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg
     1441 atatgtatac ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac
     1501 agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg
     1561 gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa
     1621 aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac
     1681 gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta
     1741 tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat
     1801 ccccctggcc agtgcacgtc tgctgtcaga taaagtcccc cgtgaacttt acccggtggt
     1861 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtttc
     1921 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     1981 taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg
     2041 tcgaccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca
     2101 aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt
     2161 acaaagtggt tggggtctca tgtctgtctt gtcttcgggg tagcggccgc ttggatccca
     2221 gaattctagg tacctcttaa ttaatggagc acaagactgg cctcatgggc cttccgctca
     2281 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaacatgg tcatagctgt
     2341 ttccttgcgt attgggcgct ctccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2401 cgggtaaagc ctggggtgcc taatgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2461 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2521 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2581 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2641 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2701 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2761 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2821 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2881 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2941 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3001 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3061 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3121 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3181 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3241 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3301 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3361 gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca gcaataaacc
     3421 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3481 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3541 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3601 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3661 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3721 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3781 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3841 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3901 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3961 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4021 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4081 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4141 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4201 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tg
//
