LOCUS       sequence-84656-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 T-Saphire noATG
FEATURES             Location/Qualifiers
     source          1..3222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..88
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(126..148)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     248..267
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     461..483
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     475..492
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     476..492
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(539..1249,1250..1252))
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(581..602)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     930..949
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1189..1210
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1350..1366)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1350..1366)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1363..1385)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1374..1390
                     /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(1398..1404,1405..1422,1423..1428))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1443..1464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1581..1598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1752..2340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1832..1851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2455..2460,2461..2477,2478..2483)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2672..3205
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gatcggcttc ccggccgcgg agttgttcgg taaattgtca caacgccgct catgacatta
       61 acctataaaa ataggcgtat cacgaggccc ttttgtctcg cgcgtttcgg tgatgacggt
      121 gaaaacctct gacacatgca gctcccggag aaggtcacag cttgtctgta agcggatgcc
      181 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
      241 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
      301 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
      361 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
      421 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
      481 acgacggcca gtgaattcga gctcggtacc tcgtagaaga caacagaggt ctcaccttct
      541 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
      601 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggatgctc aggtagtggt
      661 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
      721 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct
      781 tctgcttgtc ggccatgata tagacgttgt ggctgttgaa gttgtactcc agcttgtgcc
      841 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
      901 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
      961 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     1021 tgtggtcggg gtagcgggcg aacaccatca cgccgtagga gaaggtggtc acgagggtgg
     1081 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     1141 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     1201 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc acggtgtgag
     1261 acccgtaacg tcttcatcga atgcatctag atatcggatc ccgggcccgt cgactgcaga
     1321 ggcctgcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1381 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1441 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1501 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1561 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1621 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1681 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1741 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1801 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1861 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1921 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1981 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2041 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2101 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2161 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2221 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2281 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2341 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2401 gggattttgg tcatgaggcg cacaccgtgg aaacggatga aggcacgaac ccagttgaca
     2461 taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc
     2521 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     2581 gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     2641 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca
     2701 gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg
     2761 taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga
     2821 gttcggcgac gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt
     2881 gctccgtagt aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc
     2941 tctcgcggct tacgttctgc ccaggtttga gcagccgcgt agtgagatct atatctatga
     3001 tctcgcagtc tccggcgagc accggaggca gggcattgcc accgcgctca tcaatctcct
     3061 caagcatgag gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga
     3121 cgatcccgca gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga
     3181 tatcgaccca agtaccgcca cctaacaatt cgttcaagcc ga
//
