LOCUS       sequence-84655-e        3221 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 3221)
  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 3221)
  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 YFP noATG
FEATURES             Location/Qualifiers
     source          1..3221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..66)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     166..185
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     379..401
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     393..410
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     394..410
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(456..1166,1167..1169))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(498..519)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     847..866
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1106..1127
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1267..1283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1267..1283)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1280..1302)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1291..1307
                     /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(1315..1321,1322..1339,1340..1345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1360..1381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1498..1515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1669..2257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1749..1768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2372..2377,2378..2394,2395..2400)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2589..3122
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     join(3209..3221,1..6)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgaggccctt ttgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
       61 tcccggagaa ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      121 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      181 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      241 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      301 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      361 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
      421 tcggtacctc gagaagacaa cagaggtctc accttcttgt acagctcgtc catgccgaga
      481 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
      541 tctttgctca gggcggactg gtagctcagg tagtggttgt cgggcagcag cacggggccg
      601 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc gtcctcgatg
      661 ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc ggtgatatag
      721 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
      781 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
      841 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
      901 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcgggcgaag
      961 cactgcaggc cgtagcccag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     1021 gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     1081 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     1141 ccggtgaaca gctcctcgcc cttgctcacg gtgtgagacc cgtaaagtct tcatcgaatg
     1201 catctagata tcggatcccg ggcccgtcga ctgcagaggc ctgcatgcaa gcttggcgta
     1261 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1321 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1381 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1441 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1501 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1561 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1861 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2101 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaggcgcac
     2341 accgtggaaa cggatgaagg cacgaaccca gttgacataa gcctgttcgg ttcgtaaact
     2401 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2461 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg tacagtctat
     2521 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     2581 cagcaacgat gttacgcagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     2641 agttaggtgg ctcaagtatg ggcatcattc gcacatgtag gctcggccct gaccaagtca
     2701 aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt cggcgacgta gccacctact
     2761 cccaacatca gccggactcc gattacctcg ggaacttgct ccgtagtaag acattcatcg
     2821 cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct cgcggcttac gttctgccca
     2881 ggtttgagca gccgcgtagt gagatctata tctatgatct cgcagtctcc ggcgagcacc
     2941 ggaggcaggg cattgccacc gcgctcatca atctcctcaa gcatgaggcc aacgcgcttg
     3001 gtgcttatgt gatctacgtg caagcagatt acggtgacga tcccgcagtg gctctctata
     3061 caaagttggg catacgggaa gaagtgatgc actttgatat cgacccaagt accgccacct
     3121 aacaattcgt tcaagccgag atcggcttcc cggccgcgga gttgttcggt aaattgtcac
     3181 aacgccgctc atgacattaa cctataaaaa taggcgtatc a
//
