LOCUS       sequence-84590-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 3-4
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1267..1289
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1281..1298
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1282..1298
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1309..1328
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1309..1327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1471..1595
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1636..1941)
                     /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(1738..1757)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2283..2942)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2857..2876
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2996..3002,3003..3020,3021..3026))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(3063..3187)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3384..3401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3494..4082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3574..3593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     1021 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1081 atcccttata aatcaaaaga atagaccgag atagggttga gtggccgcta cagggcgctc
     1141 ccattcgcca ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc
     1201 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     1261 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgacgta atacgactca
     1321 ctatagggcg aattggcgga aggccgtcaa ggccacgtgt cttgtccagg cgcgccatga
     1381 gctctaaagc ttcgctcgag ctgcggccgc acggatcctc gaattcccaa gcttatctcg
     1441 acccgaagac aagacagcgg tgagacccca accactttgt acaagaaagc tgaacgagaa
     1501 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     1561 taatactgta aaacacaaca tatccagtca ctatggtcga cctgcagact ggctgtgtat
     1621 aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt tgatgtcatt
     1681 ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gaaaccggca cactggccat
     1741 atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt aaagttcacg
     1801 ggggacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc gtcgcccggg
     1861 cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct ctcttttata
     1921 ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc gtcagcaaaa gagccgttca
     1981 tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc cagcgttcgg
     2041 cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cagaccggag atattgacat
     2101 catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa tacgctgctt
     2161 catagcatac ctctttttga catacttcgg gtatacatat cagtatatat tcttataccg
     2221 caaaaatcag cgcgcaaata cgcatactgt tatctggctt ttagtaagcc ggatcctcta
     2281 gattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     2341 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     2401 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     2461 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     2521 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     2581 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     2641 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     2701 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg
     2761 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     2821 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     2881 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     2941 attttagctt ccttagctcc tgaaaatctc gacggatcct aactcaaaat ccacacatta
     3001 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgcggcccc tcgaggggcc
     3061 gccatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     3121 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct tttttgtaca
     3181 aacttgtggt ctcatgtcca atttgtcttc ggggtagcgg ccgcttggat cccagaattc
     3241 taggtacctc ttaattaatg gagcacaaga ctggcctcat gggccttccg ctcactgccc
     3301 gctttccagt cgggaaacct gtcgtgccag ctgcattaac atggtcatag ctgtttcctt
     3361 gcgtattggg cgctctccgc ttcctcgctc actgactcgc tgcgctcggt cgttcgggta
     3421 aagcctgggg tgcctaatga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac ag
//
