LOCUS       sequence-84657-e        3222 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 3222)
  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 3222)
  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 Cerulean noATG
FEATURES             Location/Qualifiers
     source          1..3222
                     /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(457..1167,1168..1170))
                     /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(499..520)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     848..867
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1107..1128
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1268..1284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1268..1284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1281..1303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1292..1308
                     /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(1316..1322,1323..1340,1341..1346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1361..1382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1499..1516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1670..2258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1750..1769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2373..2378,2379..2395,2396..2401)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2590..3123
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     join(3210..3222,1..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 gtagaagaca acagaggtct caccttcttg tacagctcgt ccatgccgag
      481 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
      541 gtctttgctc agcttggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
      601 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
      661 gttgtggcgg atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata
      721 gacgttgtcg ctgatggcgt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
      781 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
      841 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
      901 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa
      961 gcactgcacg ccccaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     1021 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     1081 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     1141 cccggtgaac agctcctcgc ccttgctcac ggtgtgagac ccgtaaagtc ttcatcgaat
     1201 gcatctagat atcggatccc gggcccgtcg actgcagagg cctgcatgca agcttggcgt
     1261 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1321 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2101 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgaggcgca
     2341 caccgtggaa acggatgaag gcacgaaccc agttgacata agcctgttcg gttcgtaaac
     2401 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     2461 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta
     2521 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     2581 gcagcaacga tgttacgcag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     2641 aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc
     2701 aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggcgacgt agccacctac
     2761 tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc
     2821 gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc
     2881 aggtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc cggcgagcac
     2941 cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt
     3001 ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat
     3061 acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc
     3121 taacaattcg ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca
     3181 caacgccgct catgacatta acctataaaa ataggcgtat ca
//
