LOCUS       sequence-84652-e        2733 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 2733)
  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 2733)
  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: LI E-F HSP-T
FEATURES             Location/Qualifiers
     source          1..2733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..154
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(192..214)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     314..333
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     527..549
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     541..558
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     542..558
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      594..842
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     primer_bind     complement(927..943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(927..943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(940..962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(975..981,982..999,1000..1005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1020..1041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1158..1175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1329..1917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1409..1428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2032..2037,2038..2054,2055..2060)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2249..2733,1..49)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca
       61 agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac gccgctcatg
      121 acattaacct ataaaaatag gcgtatcacg aggccctttt gtctcgcgcg tttcggtgat
      181 gacggtgaaa acctctgaca catgcagctc ccggagaagg tcacagcttg tctgtaagcg
      241 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
      301 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
      361 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
      421 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      481 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      541 tgtaaaacga cggccagtga attcgagctc ggtacctcga tggtctcaaa tcatatgaag
      601 atgaagatga aatatttggt gtgtcaaata aaaagcttgt gtgcttaagt ttgtgttttt
      661 ttcttggctt gttgtgttat gaatttgtgg ctttttctaa tattaaatga atgtaagatc
      721 tcattataat gaataaacaa atgtttctat aatccattgt gaatgttttg ttggatctct
      781 tctgcagcat ataactactg tatgtgctat ggtatggact atggaatatg attaaagata
      841 agtgagagag acctcgaatg catctagata tcggatcccg ggcccgtcga ctgcagaggc
      901 ctgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
      961 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1021 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1141 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1201 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1261 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1321 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1381 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1441 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1501 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1561 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1621 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1681 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1741 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1801 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1861 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1921 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1981 attttggtca tgaggcgcac accgtggaaa cggatgaagg cacgaaccca gttgacataa
     2041 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     2101 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     2161 tgtttttttg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     2221 cgatgtttga tgttatggag cagcaacgat gttacgcagc agcaacgatg ttacgcagca
     2281 gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc gcacatgtag
     2341 gctcggccct gaccaagtca aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt
     2401 cggcgacgta gccacctact cccaacatca gccggactcc gattacctcg ggaacttgct
     2461 ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct
     2521 cgcggcttac gttctgccca ggtttgagca gccgcgtagt gagatctata tctatgatct
     2581 cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc gcgctcatca atctcctcaa
     2641 gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg caagcagatt acggtgacga
     2701 tcccgcagtg gctctctata caaagttggg cat
//
