LOCUS       sequence-84643-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 YFP
FEATURES             Location/Qualifiers
     source          1..3256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     348..366
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(404..426)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     526..545
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     739..761
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     753..770
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     754..770
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(856..858,859..861,862..1572)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(901..922)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1162..1181)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1509..1530
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1662..1678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1662..1678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1675..1697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1686..1702
                     /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(1710..1716,1717..1734,1735..1740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1755..1776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1893..1910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2064..2652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2144..2163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2767..2772,2773..2789,2790..2795)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(2984..3256,1..261)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gcggcttacg ttctgcccag gtttgagcag ccgcgtagtg agatctatat ctatgatctc
       61 gcagtctccg gcgagcaccg gaggcagggc attgccaccg cgctcatcaa tctcctcaag
      121 catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta cggtgacgat
      181 cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca ctttgatatc
      241 gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc ggccgcggag
      301 ttgttcggta aattgtcaca acgccgctca tgacattaac ctataaaaat aggcgtatca
      361 cgaggccctt ttgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      421 tcccggagaa ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      481 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      541 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      601 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      661 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      721 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
      781 tcggtacctc gcgaatgcat ctagatatcg gatcccgggc ccgtcgactg cagaggtacg
      841 ggtctcatct gaacaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      901 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      961 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1021 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     1081 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1141 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1201 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1261 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1321 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1381 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1441 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     1501 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1561 gagctgtaca agggtggagg aggttctgga ggcggtggaa gtggtggcgg aggtagcggc
     1621 acctgagacc tctgcctgca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     1681 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1741 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1801 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1981 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2701 aactcacgtt aagggatttt ggtcatgagg cgcacaccgt ggaaacggat gaaggcacga
     2761 acccagttga cataagcctg ttcggttcgt aaactgtaat gcaagtagcg tatgcgctca
     2821 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
     2881 atggcttgtt atgactgttt ttttgtacag tctatgcctc gggcatccaa gcagcaagcg
     2941 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagcaa
     3001 cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggtggctcaa gtatgggcat
     3061 cattcgcaca tgtaggctcg gccctgacca agtcaaatcc atgcgggctg ctcttgatct
     3121 tttcggtcgt gagttcggcg acgtagccac ctactcccaa catcagccgg actccgatta
     3181 cctcgggaac ttgctccgta gtaagacatt catcgcgctt gctgccttcg accaagaagc
     3241 ggttgttggc gctctc
//
