LOCUS       sequence-84659-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 mOrange noATG
FEATURES             Location/Qualifiers
     source          1..3213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     466..484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(522..544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     644..663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(743..1741)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAGLCSRRARDPISRCIRRRQQRSHLLVQLVHAAGGV
                     AALGAFVLFHDGVVLVVGGDVQLDADDVGAGQLHGLLGLVGGLDLGGVVAAVLQLQPHL
                     DLALQGAVLGVHPLGGGLPAHGFLLHYGAVGGEVGAAQLHLVDELAVLQGGVLGHGHHA
                     AVLEVHHALPLEALGEGQLEVVGDVGGVLHVGLGAVGELRGQDVPGEGQGATLGHLQLS
                     GLKALVGAALALALDLELVAVHRALHAHLEAHELLDDGHVILLALAHGVRPVKSSSRYR
                     ARIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     857..879
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     871..888
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     872..888
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             935..1639
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGF
                     QTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1737..1753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1737..1753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1750..1772)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1761..1777
                     /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(1785..1791,1792..1809,1810..1815))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1830..1851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1968..1985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2139..2727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2219..2238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2842..2847,2848..2864,2865..2870)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3059..3213,1..379)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cgacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
       61 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
      121 ggcttacgtt ctgcccaggt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
      181 agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
      241 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
      301 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
      361 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
      421 gttcggtaaa ttgtcacaac gccgctcatg acattaacct ataaaaatag gcgtatcacg
      481 aggccctttt gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
      541 ccggagaagg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      601 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      661 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      721 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      781 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      841 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
      901 ggtacctcga tgaagacttt acgggtctca caccgtgagc aagggcgagg agaataacat
      961 ggccatcatc aaggagttca tgcgcttcaa ggtgcgcatg gagggctctg tgaacggcca
     1021 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggctttc agaccgctaa
     1081 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1141 cacctacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttcaagct
     1201 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1261 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1321 cggcaccaac ttcccctccg acggccccgt aatgcagaag aaaaccatgg gctgggaggc
     1381 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct
     1441 gaagctgaag gacggcggcc actacacctc cgaggtcaag accacctaca aggccaagaa
     1501 gcccgtgcag ctgcccggcg cctacatcgt cggcatcaag ttggacatca cctcccacaa
     1561 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1621 catggacgag ctgtacaaga aggtgagacc tctgttgtct tctacgaatg catctagata
     1681 tcggatcccg ggcccgtcga ctgcagaggc ctgcatgcaa gcttggcgta atcatggtca
     1741 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1801 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1981 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2041 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaggcgcac accgtggaaa
     2821 cggatgaagg cacgaaccca gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag
     2881 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     2941 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg tacagtctat gcctcgggca
     3001 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     3061 gttacgcagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggtgg
     3121 ctcaagtatg ggcatcattc gcacatgtag gctcggccct gaccaagtca aatccatgcg
     3181 ggctgctctt gatcttttcg gtcgtgagtt cgg
//
