LOCUS       sequence-84658-e        3213 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 3213)
  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 3213)
  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 C-D mCherry noATG
FEATURES             Location/Qualifiers
     source          1..3213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..54
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(92..114)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     214..233
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     427..449
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     441..458
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     442..458
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(505..1209)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     1049..1067
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1307..1323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1307..1323)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1320..1342)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1331..1347
                     /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(1355..1361,1362..1379,1380..1385))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1400..1421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1538..1555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1709..2297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1789..1808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2412..2417,2418..2434,2435..2440)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2629..3162
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ttgtcacaac gccgctcatg acattaacct ataaaaatag gcgtatcacg aggccctttt
       61 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagaagg
      121 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
      181 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
      241 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
      301 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
      361 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      421 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc ggtacctcgt
      481 agaagacaac agaggtctca ccttcttgta cagctcgtcc atgccgccgg tggagtggcg
      541 gccctcggcg cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc
      601 caacttgatg ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt
      661 ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct gcttgatctc
      721 gcccttcagg gcgccgtcct cggggtacat ccgctcggag gaggcctccc agcccatggt
      781 tttcttctgc attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta
      841 gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc
      901 gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttca agtagtcggg
      961 gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac atgaactgag gggacaggat
     1021 gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc
     1081 ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcacggagcc
     1141 ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat cctcctcgcc
     1201 cttgctcacg gtgtgagacc cgtaaagtct tcatcgaatg catctagata tcggatcccg
     1261 ggcccgtcga ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc
     1321 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1381 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1441 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1501 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1561 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1621 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1681 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1741 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1801 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1861 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1921 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1981 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2041 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2101 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2161 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2221 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2281 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2341 acgaaaactc acgttaaggg attttggtca tgaggcgcac accgtggaaa cggatgaagg
     2401 cacgaaccca gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc
     2461 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     2521 ttttcatggc ttgttatgac tgtttttttg tacagtctat gcctcgggca tccaagcagc
     2581 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     2641 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg
     2701 ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg ggctgctctt
     2761 gatcttttcg gtcgtgagtt cggcgacgta gccacctact cccaacatca gccggactcc
     2821 gattacctcg ggaacttgct ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa
     2881 gaagcggttg ttggcgctct cgcggcttac gttctgccca ggtttgagca gccgcgtagt
     2941 gagatctata tctatgatct cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc
     3001 gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg
     3061 caagcagatt acggtgacga tcccgcagtg gctctctata caaagttggg catacgggaa
     3121 gaagtgatgc actttgatat cgacccaagt accgccacct aacaattcgt tcaagccgag
     3181 atcggcttcc cggccgcgga gttgttcggt aaa
//
