LOCUS       sequence-84638-e        3196 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 3196)
  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 3196)
  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 c1
FEATURES             Location/Qualifiers
     source          1..3196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     223..241
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(279..301)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     401..420
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     614..636
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     628..645
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     629..645
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             727..1434
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(872..890)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1477..1493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1477..1493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1490..1512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1501..1517
                     /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(1525..1531,1532..1549,1550..1555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1570..1591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1708..1725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1879..2467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1959..1978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2582..2587,2588..2604,2605..2610)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2799..3196,1..136)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca gattacggtg acgatcccgc
       61 agtggctctc tatacaaagt tgggcatacg ggaagaagtg atgcactttg atatcgaccc
      121 aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttcccggccg cggagttgtt
      181 cggtaaattg tcacaacgcc gctcatgaca ttaacctata aaaataggcg tatcacgagg
      241 cccttttgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      301 gagaaggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      361 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      421 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      481 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      541 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
      601 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt
      661 acctcgcgaa tgcatctaga tatcggatcc cgggcccgtc gactgcagag gtacgggtct
      721 cacaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      781 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      841 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      901 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      961 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1021 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1081 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1141 cccgtaatgc agaagaaaac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1201 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1261 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1321 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1381 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagaaggtg
     1441 agacctctgc ctgcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1501 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1561 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1621 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1681 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1741 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1801 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2401 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2521 acgttaaggg attttggtca tgaggcgcac accgtggaaa cggatgaagg cacgaaccca
     2581 gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa
     2641 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     2701 ttgttatgac tgtttttttg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     2761 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agcaacgatg
     2821 ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc
     2881 gcacatgtag gctcggccct gaccaagtca aatccatgcg ggctgctctt gatcttttcg
     2941 gtcgtgagtt cggcgacgta gccacctact cccaacatca gccggactcc gattacctcg
     3001 ggaacttgct ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa gaagcggttg
     3061 ttggcgctct cgcggcttac gttctgccca ggtttgagca gccgcgtagt gagatctata
     3121 tctatgatct cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc gcgctcatca
     3181 atctcctcaa gcatga
//
