LOCUS       sequence-84592-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 5-6
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..118
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(119..187,188..979)
                     /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(337..356)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1150..1738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1639..1658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1831..1848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2045..2169
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2206..2211,2212..2229,2230..2236)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2290..2949
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2356..2375)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3291..3596
                     /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     3475..3494
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3637..3761)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3904..3923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3905..3923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3934..3951)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3934..3950)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3943..3965)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
       61 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      121 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      181 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      241 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      301 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      361 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      421 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      481 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      541 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      601 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      661 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      721 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      781 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggttctcg
      841 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      901 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      961 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1261 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcattaggc
     1801 accccaggct ttacccgaac gaccgagcgc agcgagtcag tgagcgagga agcggagagc
     1861 gcccaatacg caaggaaaca gctatgacca tgttaatgca gctggcacga caggtttccc
     1921 gactggaaag cgggcagtga gcggaaggcc catgaggcca gtcttgtgct ccattaatta
     1981 agaggtacct agaattctgg gatccaagcg gccgctaccc cgaagacaat gaggcggtga
     2041 gaccacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat
     2101 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
     2161 gtcactatgg cggcccctcg aggggccgca ttaggcaccc caggctttac actttatgct
     2221 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
     2281 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     2341 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     2401 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     2461 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     2521 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     2581 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     2641 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     2701 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     2761 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     2821 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     2881 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     2941 ggggcgtaat ctagaggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
     3001 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
     3061 ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
     3121 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
     3181 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
     3241 gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta
     3301 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     3361 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     3421 ataaagtccc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     3481 tgaccaccga tatggccagt gtgccggttt ccgttatcgg ggaagaagtg gctgatctca
     3541 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     3601 caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt
     3661 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     3721 tcattttacg tttctcgttc agctttcttg tacaaagtgg ttggggtctc atgtctctgt
     3781 tgtcttcggg tcgagataag cttgggaatt cgaggatccg tgcggccgca gctcgagcga
     3841 agctttagag ctcatggcgc gcctggacaa gacacgtggc cttgacggcc ttccgccaat
     3901 tcgccctata gtgagtcgta ttacgtcgcg ctcactggcc gtcgttttac aacgtcgtga
     3961 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4021 ctggcgtaat agcgaagagg cccgcaccga aacgcccttc ccaacagttg cgcagcctga
     4081 atggcgaatg ggagcgccct gtagcggcca ctcaacccta tctcggtcta ttcttttgat
     4141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4201 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tt
//
