LOCUS       sequence-84654-e        3222 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 3222)
  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 3222)
  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 GFP  noATG
FEATURES             Location/Qualifiers
     source          1..3222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..108
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(146..168)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     268..287
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(367..1374)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAGLCSRRARDPISRCIRRRQQRSHLLVQLVHAVSDP
                     GGGHELQQDHVIALLVGVFAQGGLGAQVVVVGQQHGAVADGGVLLVVVGELHAAVLDVV
                     ADLEVHLDAVLLLVGHDIDVVAVVVVLQLVPQDVAVLLEVDALQLDAVHQGVALELHLG
                     AGLVVAVVLEEDGALLDVAFGHGGLEEVVLLHVVGVAAEALHAVGEGGHEGGPGHGQLA
                     GGADELQGQLAVGGIALALAGHAELVAVYVAVQLDQDGHHPGEQLLALAHGVRPVKSSS
                     RYRARIHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPNSCAA"
     primer_bind     481..503
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     495..512
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     496..512
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(559..561,562..1272)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(601..622)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(862..881)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1209..1230
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1370..1386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1370..1386)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1383..1405)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1394..1410
                     /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(1418..1424,1425..1442,1443..1448))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1463..1484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1601..1618)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1772..2360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1852..1871)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2475..2480,2481..2497,2498..2503)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2692..3222,1..3)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 taacaattcg ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca
       61 caacgccgct catgacatta acctataaaa ataggcgtat cacgaggccc ttttgtctcg
      121 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag aaggtcacag
      181 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
      241 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
      301 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
      361 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
      421 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
      481 cccagtcacg acgttgtaaa acgacggcca gtgaattcga gctcggtacc tcgatgaaga
      541 ctttacgggt ctcacaccgt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      721 gtgccctggc ccaccctcgt gaccaccttc acctacggcg tgcagtgctt cagccgctac
      781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1201 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca cggcatggac
     1261 gagctgtaca agaaggtgag acctctgttg tcttctacga atgcatctag atatcggatc
     1321 ccgggcccgt cgactgcaga ggcctgcatg caagcttggc gtaatcatgg tcatagctgt
     1381 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1441 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1501 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1561 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1621 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1681 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1741 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1801 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1861 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1921 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1981 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2041 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2101 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2161 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2221 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2281 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2341 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2401 ggaacgaaaa ctcacgttaa gggattttgg tcatgaggcg cacaccgtgg aaacggatga
     2461 aggcacgaac ccagttgaca taagcctgtt cggttcgtaa actgtaatgc aagtagcgta
     2521 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     2581 cggttttcat ggcttgttat gactgttttt ttgtacagtc tatgcctcgg gcatccaagc
     2641 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
     2701 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt
     2761 atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct
     2821 cttgatcttt tcggtcgtga gttcggcgac gtagccacct actcccaaca tcagccggac
     2881 tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc tgccttcgac
     2941 caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaggtttga gcagccgcgt
     3001 agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca gggcattgcc
     3061 accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta tgtgatctac
     3121 gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt gggcatacgg
     3181 gaagaagtga tgcactttga tatcgaccca agtaccgcca cc
//
