LOCUS       sequence-84587-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 1-2
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(155..223,224..1015)
                     /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(373..392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1186..1774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1675..1694
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1867..1884
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2081..2205
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2242..2247,2248..2265,2266..2272)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2326..2985
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2392..2411)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3327..3632
                     /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     3511..3530
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3673..3797)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3940..3959)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3941..3959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3970..3987)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3970..3986)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3979..4001)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
       61 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      121 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      181 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
      241 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
      301 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
      361 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
      421 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
      481 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
      541 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
      601 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
      661 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
      721 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
      781 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
      841 tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt atcattgcag cactggggcc
      901 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
      961 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1021 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1081 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1141 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1201 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1261 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1321 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1381 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1441 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1501 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1561 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1621 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1681 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1741 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1801 gttcctggcc ttttgctggc cttttgctca ttaggcaccc caggctttac ccgaacgacc
     1861 gagcgcagcg agtcagtgag cgaggaagcg gagagcgccc aatacgcaag gaaacagcta
     1921 tgaccatgtt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgg
     1981 aaggcccatg aggccagtct tgtgctccat taattaagag gtacctagaa ttctgggatc
     2041 caagcggccg ctaccccgaa gacaatacgg acatgagacc acaagtttgt acaaaaaagc
     2101 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa
     2161 acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc ccctcgaggg
     2221 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     2281 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     2341 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     2401 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     2461 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     2521 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     2581 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     2641 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     2701 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     2761 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2821 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2881 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     2941 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatctag aggatccggc
     3001 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     3061 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
     3121 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     3181 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     3241 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     3301 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     3361 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     3421 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtcccccgt gaactttacc
     3481 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     3541 cggtttccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     3601 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     3661 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     3721 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     3781 ttcttgtaca aagtggttgg ggtctcaccg cgggcttgtc ttcgggtcga gataagcttg
     3841 ggaattcgag gatccgtgcg gccgcagctc gagcgaagct ttagagctca tggcgcgcct
     3901 ggacaagaca cgtggccttg acggccttcc gccaattcgc cctatagtga gtcgtattac
     3961 gtcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4021 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4081 caccgaaacg cccttcccaa cagttgcgca gcctgaatgg cgaatgggag cgccctgtag
     4141 cggccactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4201 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt ta
//
