LOCUS       sequence-84593-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 R 5-6
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     288..310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     302..319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     303..319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..349
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        330..348
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    492..616
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(657..962)
                     /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     complement(759..778)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1304..1963)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1878..1897
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2017..2023,2024..2041,2042..2047))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2084..2208)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2405..2422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2515..3103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2595..2614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3274..4065,4066..4134))
                     /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     3897..3916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4135..4239)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
       61 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
      121 aatagaccga gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg
      181 cgcaactgtt gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      241 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      301 ttgtaaaacg acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg
      361 aaggccgtca aggccacgtg tcttgtccag gcgcgccatg agctctaaag cttcgctcga
      421 gctgcggccg cacggatcct cgaattccca agcttatctc gacccgaaga caacagagcg
      481 gtgagacccc aaccactttg tacaagaaag ctgaacgaga aacgtaaaat gatataaata
      541 tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac
      601 atatccagtc actatggtcg acctgcagac tggctgtgta taagggagcc tgacatttat
      661 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca
      721 gccacttctt ccccgataac ggaaaccggc acactggcca tatcggtggt catcatgcgc
      781 cagctttcat ccccgatatg caccaccggg taaagttcac gggggacttt atctgacagc
      841 agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc
      901 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc
      961 atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac
     1021 ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc
     1081 attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac
     1141 tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg
     1201 acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat
     1261 acgcatactg ttatctggct tttagtaagc cggatcctct agattacgcc ccgccctgcc
     1321 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     1381 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     1441 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     1501 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
     1561 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     1621 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     1681 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     1741 tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     1801 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     1861 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     1921 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     1981 ctgaaaatct cgacggatcc taactcaaaa tccacacatt atacgagccg gaagcataaa
     2041 gtgtaaagcc tggggtgcct aatgcggccc ctcgaggggc cgccatagtg actggatatg
     2101 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     2161 atttatatca ttttacgttt ctcgttcagc ttttttgtac aaacttgtgg tctcatgtcc
     2221 tcattgtctt cggggtagcg gccgcttgga tcccagaatt ctaggtacct cttaattaat
     2281 ggagcacaag actggcctca tgggccttcc gctcactgcc cgctttccag tcgggaaacc
     2341 tgtcgtgcca gctgcattaa catggtcata gctgtttcct tgcgtattgg gcgctctccg
     2401 cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg
     2461 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2521 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2581 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2641 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2701 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2761 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2821 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2881 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2941 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3001 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3061 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3121 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3181 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3241 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3301 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3361 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc
     3421 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3481 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3541 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3601 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3661 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3721 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3781 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3841 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3901 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3961 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4021 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4081 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4141 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4201 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc ga
//
