LOCUS       sequence-84622-e        5495 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 5495)
  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 5495)
  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 1-2 RNAi
FEATURES             Location/Qualifiers
     source          1..5495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..18
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(1491..1496,1497..1514,1515..1521)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1575..2234
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1641..1660)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2576..2881
                     /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     2760..2779
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(3317..3336)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3318..3336)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3347..3364)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3347..3363)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3356..3378)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        3679..3783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3784..3852,3853..4644)
                     /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     complement(4002..4021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4815..5403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5304..5323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgagtcag tgagcgagga agcggagagc gcccaatacg caaggaaaca gctatgacca
       61 tgttaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcggaaggcc
      121 catgaggcca gtcttgtgct ccattaatta agaggtacct agaattctgg gatccaagcg
      181 gccgctaccc cgaagacaat acggcgggga gagaggattt tgaggaaata attaattgaa
      241 ttacttgtat tattgataaa gtaatttaga taagttgtta gtacaactta ttgcaactat
      301 ggatagacaa aaatcactta ttttaagggg agaagtatat aacaacttat tataaaattt
      361 ctgagcagcc ggcctcctcc aatcaatcat agagagtgga gcccattctg gaaaaccaag
      421 gaacccccaa cttgcactgg tgcggtggcc caattcaaaa aaaacggggc ccagcaaagt
      481 aacgccgttg taacgattta tcaatccaaa ctcaaaaggc ggcgagatgc gtttaactcc
      541 gtgaaattaa caaaccgcca acaacttgca atttgcaact accgtttcca agaagaactc
      601 aaccacacaa cgtatcctat cccaaaccac acgcaactag tgacgcgtca taaggacacg
      661 tgtcacaatt tgactggtta ataatttcac cgcttttgct ataaattacc tccaatcccc
      721 ttagcttctt cacaattcag ttcccaaccc taacaattct tgttcatatc gcttctctct
      781 actttcaagg tatgatccaa tttctctctt ctttctctgt aatcctttcg ttgagttttg
      841 tttccgatca atcataggta gttttcttgt ttcgaagcat gagatctagg aattttttgt
      901 gattttccaa aattgagatc ggtttgaaat tgaattttac agcttgaatc tcagatcttg
      961 ttttatcaat gttttcgatg gcttgcgatg tagatctatg ataattgtgg ttcagttttg
     1021 ttaggaatcg atttcggttt agcaattgca gattaattag ggtttccaat tgaattcttc
     1081 agatccgtta tggaattatg tcaaataatt tattcaaatt ggaaattatt gttagatcca
     1141 ctcttaatct gtttgatcca agcttcaatt agggttttca cttgtttcaa tttcttgtta
     1201 tggattctga tttatctgtt gatgttagat ccactcttaa tctgtttgat ccaagcgtta
     1261 attagggttt tcacttgttt caatttcgtg tgttggattc tgatttatct gttgttgatg
     1321 tgattacagt ctgcacctga gaccaaagct gaacgagaaa cgtaaaatga tataaatatc
     1381 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     1441 atccagtcac tatggcggcc cctcgagggg ccgcattagg caccccaggc tttacacttt
     1501 atgcttccgg ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag
     1561 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     1621 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     1681 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     1741 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
     1801 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     1861 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     1921 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     1981 aagggtttat tgagaatatg tttttcgtgt cagccaatcc ctgggtgagt ttcaccagtt
     2041 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     2101 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     2161 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     2221 agggcggggc gtaatctaga ggatccggct tactaaaagc cagataacag tatgcgtatt
     2281 tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa
     2341 aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt
     2401 gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa
     2461 gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc
     2521 gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca
     2581 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
     2641 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
     2701 gtcagataaa gtcccccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
     2761 catgatgacc accgatatgg ccagtgtgcc ggtttccgtt atcggggaag aagtggctga
     2821 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
     2881 aatgtcaggc tcccttatac acagccagtc tgcaggtcga cggtctcagg tgaatccggc
     2941 catgctagag tccgcaaaaa tcaccagtct ctctctacaa atctatctct ctctattttt
     3001 ctccagaata atgtgtgagt agttcccaga taagggaatt agggttctta tagggtttcg
     3061 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     3121 tcaataaaat ttctaattcc taaaaccaaa atccagtgac cttgagtgac ctaagctagc
     3181 ctagtgtcgg gcttgtcttc gggtcgagat aagcttggga attcgaggat ccgtgcggcc
     3241 gcagctcgag cgaagcttta gagctcatgg cgcgcctgga caagacacgt ggccttgacg
     3301 gccttccgcc aattcgccct atagtgagtc gtattacgtc gcgctcactg gccgtcgttt
     3361 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3421 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgaaacgccc ttcccaacag
     3481 ttgcgcagcc tgaatggcga atgggagcgc cctgtagcgg ccactcaacc ctatctcggt
     3541 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3601 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3661 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3721 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3781 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3841 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3901 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3961 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4021 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4081 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4141 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4201 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4261 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4321 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4381 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4441 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggt
     4501 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4561 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4621 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4681 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4741 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4801 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4861 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4921 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4981 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5041 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5101 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5161 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5221 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5281 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5341 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5401 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcat
     5461 taggcacccc aggctttacc cgaacgaccg agcgc
//
