LOCUS       sequence-84671-e        3414 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 3414)
  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 3414)
  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 destab GFP c1
FEATURES             Location/Qualifiers
     source          1..3414
                     /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..1942)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAGLCSRRARDPISRCIRRSHTMGLINIINNPLKNSL
                     KWIINYRTGITLLSLVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     KVRPTRYRARIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA
                     "
     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             complement(join(923..1042,1043..1048,1049..1759,
                     1760..1762))
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV"
     primer_bind     complement(1091..1112)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1440..1459
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1699..1720
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1938..1954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1938..1954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1951..1973)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1962..1978
                     /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(1986..1992,1993..2010,2011..2016))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2031..2052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2169..2186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2340..2928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2420..2439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3043..3048,3049..3065,3066..3071)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3260..3414,1..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 ggtacctcgt aggtctcacc ttcacattga tcctagcaga agcacaggct gcagggtgac
      961 ggtccatccc gctctcctgg gcacaagaca tgggcagcgt gccatcatcc tgctcctcca
     1021 cctccggcgg gaagccatgg ctaagcttct tgtacagctc gtccatgccg agagtgatcc
     1081 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     1141 tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     1201 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     1261 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     1321 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     1381 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     1441 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     1501 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     1561 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     1621 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     1681 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     1741 acagctcctc gcccttgctc actagggata acagggtaat accggtccta taattaataa
     1801 tccactttaa agaattctta agtggattat taattatatt aattaagccc atggtgtgag
     1861 acctacgaat gcatctagat atcggatccc gggcccgtcg actgcagagg cctgcatgca
     1921 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1981 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2041 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2101 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2161 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2221 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2281 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2341 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2401 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2461 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2521 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2581 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2641 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2701 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2761 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2821 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2881 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2941 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3001 atgaggcgca caccgtggaa acggatgaag gcacgaaccc agttgacata agcctgttcg
     3061 gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
     3121 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     3181 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
     3241 atgttatgga gcagcaacga tgttacgcag cagcaacgat gttacgcagc agggcagtcg
     3301 ccctaaaaca aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc
     3361 tgaccaagtc aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcgg
//
