LOCUS       sequence-84642-e        3256 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 3256)
  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 3256)
  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 B-C GFP
FEATURES             Location/Qualifiers
     source          1..3256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..65)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     165..184
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     378..400
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     392..409
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     393..409
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(495..497,498..500,501..1211)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(540..561)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(801..820)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1148..1169
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1301..1317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1301..1317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1314..1336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1325..1341
                     /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(1349..1355,1356..1373,1374..1379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1394..1415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1532..1549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1703..2291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1783..1802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2406..2411,2412..2428,2429..2434)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2623..3156
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     join(3243..3256,1..5)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaggcccttt tgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
       61 cccggagaag gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      121 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      181 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      241 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      301 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      361 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
      421 cggtacctcg cgaatgcatc tagatatcgg atcccgggcc cgtcgactgc agaggtacgg
      481 gtctcatctg aacaatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      541 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      601 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      661 tgccctggcc caccctcgtg accaccttca cctacggcgt gcagtgcttc agccgctacc
      721 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      781 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      841 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
      901 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
      961 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1021 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1081 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1141 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac ggcatggacg
     1201 agctgtacaa gggtggagga ggttctggag gcggtggaag tggtggcgga ggtagcggca
     1261 cctgagacct ctgcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1321 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1381 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1441 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1501 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1561 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1621 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1681 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2101 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2341 actcacgtta agggattttg gtcatgaggc gcacaccgtg gaaacggatg aaggcacgaa
     2401 cccagttgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac
     2461 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     2521 tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag cagcaagcgc
     2581 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagcaac
     2641 gatgttacgc agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc
     2701 attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt
     2761 ttcggtcgtg agttcggcga cgtagccacc tactcccaac atcagccgga ctccgattac
     2821 ctcgggaact tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg
     2881 gttgttggcg ctctcgcggc ttacgttctg cccaggtttg agcagccgcg tagtgagatc
     2941 tatatctatg atctcgcagt ctccggcgag caccggaggc agggcattgc caccgcgctc
     3001 atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca
     3061 gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg
     3121 atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatcggc
     3181 ttcccggccg cggagttgtt cggtaaattg tcacaacgcc gctcatgaca ttaacctata
     3241 aaaataggcg tatcac
//
