LOCUS       sequence-84586-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 1-2
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        60..164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(165..233,234..1025)
                     /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(383..402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1196..1784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1685..1704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1877..1894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2091..2215
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2252..2257,2258..2275,2276..2282)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2336..2995
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2402..2421)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3337..3642
                     /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     3521..3540
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3683..3807)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3950..3969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3951..3969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3980..3997)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3980..3996)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3989..4011)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
       61 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      121 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      181 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      241 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
      301 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      361 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
      421 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
      481 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
      541 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
      601 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
      661 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
      721 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
      781 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
      841 gctggtttat tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt atcattgcag
      901 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
      961 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1021 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1081 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1141 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1201 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1261 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1321 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1381 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1441 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1501 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1561 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1621 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1681 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1741 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1801 cctttttacg gttcctggcc ttttgctggc cttttgctca ttaggcaccc caggctttac
     1861 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gagagcgccc aatacgcaag
     1921 gaaacagcta tgaccatgtt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1981 cagtgagcgg aaggcccatg aggccagtct tgtgctccat taattaagag gtacctagaa
     2041 ttctgggatc caagcggccg ctaccccgaa gacaatacgg cggtgagacc acaagtttgt
     2101 acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
     2161 tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc
     2221 ccctcgaggg gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa
     2281 tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga
     2341 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     2401 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     2461 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     2521 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     2581 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     2641 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     2701 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     2761 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     2821 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     2881 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     2941 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatctag
     3001 aggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg
     3061 tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag
     3121 cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg
     3181 tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg
     3241 aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga
     3301 acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat
     3361 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc
     3421 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtcccccgt
     3481 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     3541 gccagtgtgc cggtttccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     3601 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     3661 cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt
     3721 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
     3781 tcgttcagct ttcttgtaca aagtggttgg ggtctcatgt cgggcttgtc ttcgggtcga
     3841 gataagcttg ggaattcgag gatccgtgcg gccgcagctc gagcgaagct ttagagctca
     3901 tggcgcgcct ggacaagaca cgtggccttg acggccttcc gccaattcgc cctatagtga
     3961 gtcgtattac gtcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4021 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4081 aagaggcccg caccgaaacg cccttcccaa cagttgcgca gcctgaatgg cgaatgggag
     4141 cgccctgtag cggccactca accctatctc ggtctattct tttgatttat aagggatttt
     4201 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta ac
//
