LOCUS       sequence-84644-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 T-Saphire
FEATURES             Location/Qualifiers
     source          1..3256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     524..542
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(580..602)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     702..721
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     915..937
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     929..946
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     930..946
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1032..1034,1035..1037,1038..1748)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1077..1098)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1338..1357)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1685..1706
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1838..1854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1838..1854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1851..1873)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1862..1878
                     /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(1886..1892,1893..1910,1911..1916))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1931..1952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2069..2086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2240..2828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2320..2339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2943..2948,2949..2965,2966..2971)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3160..3256,1..437)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gccctgacca agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggcg
       61 acgtagccac ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta
      121 gtaagacatt catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg
      181 cttacgttct gcccaggttt gagcagccgc gtagtgagat ctatatctat gatctcgcag
      241 tctccggcga gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg
      301 aggccaacgc gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg
      361 cagtggctct ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc
      421 caagtaccgc cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt
      481 tcggtaaatt gtcacaacgc cgctcatgac attaacctat aaaaataggc gtatcacgag
      541 gcccttttgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      601 ggagaaggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      661 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      721 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
      781 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      841 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
      901 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgagctcgg
      961 tacctcgcga atgcatctag atatcggatc ccgggcccgt cgactgcaga ggtacgggtc
     1021 tcatctgaac aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1081 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1141 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1201 cctggcccac cctcgtgacc accttctcct acggcgtgat ggtgttcgcc cgctaccccg
     1261 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1321 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1381 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1441 tcctggggca caagctggag tacaacttca acagccacaa cgtctatatc atggccgaca
     1501 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1561 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1621 ccgacaacca ctacctgagc atccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1681 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1741 tgtacaaggg tggaggaggt tctggaggcg gtggaagtgg tggcggaggt agcggcacct
     1801 gagacctctg cctgcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1861 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1921 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1981 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2881 cacgttaagg gattttggtc atgaggcgca caccgtggaa acggatgaag gcacgaaccc
     2941 agttgacata agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca
     3001 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3061 cttgttatga ctgttttttt gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     3121 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagcaacgat
     3181 gttacgcagc agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt
     3241 cgcacatgta ggctcg
//
