LOCUS       sequence-84633-e        3252 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 3252)
  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 3252)
  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 D-E Cerulean + linker c5
FEATURES             Location/Qualifiers
     source          1..3252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..600
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     687..705
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(743..765)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     865..884
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1078..1100
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1092..1109
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1093..1109
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1238..1240,1241..1954)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1280..1301)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1541..1560)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1888..1909
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1997..2013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1997..2013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2010..2032)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2021..2037
                     /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(2045..2051,2052..2069,2070..2075))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2090..2111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2228..2245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2399..2987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2479..2498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3102..3107,3108..3124,3125..3130)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
       61 gcaacgatgt tacgcagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
      121 ttaggtggct caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa
      181 tccatgcggg ctgctcttga tcttttcggt cgtgagttcg gcgacgtagc cacctactcc
      241 caacatcagc cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg
      301 cttgctgcct tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccagg
      361 tttgagcagc cgcgtagtga gatctatatc tatgatctcg cagtctccgg cgagcaccgg
      421 aggcagggca ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt
      481 gcttatgtga tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca
      541 aagttgggca tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa
      601 caattcgttc aagccgagat cggcttcccg gccgcggagt tgttcggtaa attgtcacaa
      661 cgccgctcat gacattaacc tataaaaata ggcgtatcac gaggcccttt tgtctcgcgc
      721 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagaag gtcacagctt
      781 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      841 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      901 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
      961 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     1021 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     1081 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacctcg cgaatgcatc
     1141 tagatatcgg atcccgggcc cgtcgactgc agaggtacgg gtctcaaagg gaggtggagg
     1201 aggttctgga ggcggtggaa gtggtggcgg aggtagcgtg agcaagggcg aggagctgtt
     1261 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1321 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1381 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt
     1441 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1501 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1561 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1621 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacg ccatcagcga
     1681 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1741 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1801 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     1861 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1921 gatcactctc ggcatggacg agctgtacaa gtaaaatctg agacctctgc ctgcatgcaa
     1981 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2041 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2101 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2161 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2221 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2281 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2341 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2401 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2461 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2521 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2581 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2641 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2701 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2761 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2821 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2881 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2941 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3001 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3061 tgaggcgcac accgtggaaa cggatgaagg cacgaaccca gttgacataa gcctgttcgg
     3121 ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     3181 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     3241 tacagtctat gc
//
