LOCUS       sequence-84631-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 YFP + linker c1
FEATURES             Location/Qualifiers
     source          1..3252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     373..391
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(429..451)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     551..570
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     764..786
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     778..795
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     779..795
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(924..926,927..1640)
                     /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(966..987)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1227..1246)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1574..1595
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1683..1699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1683..1699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1696..1718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1707..1723
                     /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(1731..1737,1738..1755,1756..1761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1776..1797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1914..1931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2085..2673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2165..2184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2788..2793,2794..2810,2811..2816)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(3005..3252,1..286)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg cccaggtttg agcagccgcg
       61 tagtgagatc tatatctatg atctcgcagt ctccggcgag caccggaggc agggcattgc
      121 caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta
      181 cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg
      241 ggaagaagtg atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc
      301 cgagatcggc ttcccggccg cggagttgtt cggtaaattg tcacaacgcc gctcatgaca
      361 ttaacctata aaaataggcg tatcacgagg cccttttgtc tcgcgcgttt cggtgatgac
      421 ggtgaaaacc tctgacacat gcagctcccg gagaaggtca cagcttgtct gtaagcggat
      481 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
      541 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
      601 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
      661 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
      721 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
      781 aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa tgcatctaga tatcggatcc
      841 cgggcccgtc gactgcagag gtacgggtct caaagggagg tggaggaggt tctggaggcg
      901 gtggaagtgg tggcggaggt agcgtgagca agggcgagga gctgttcacc ggggtggtgc
      961 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1021 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     1081 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc
     1141 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1201 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1261 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1321 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1381 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     1441 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1501 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     1561 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1621 tggacgagct gtacaagtaa aatctgagac ctctgcctgc atgcaagctt ggcgtaatca
     1681 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1741 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1801 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1861 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1921 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1981 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2041 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2101 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2161 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2221 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2281 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2341 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2401 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2461 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2521 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2581 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2641 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2701 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag gcgcacaccg
     2761 tggaaacgga tgaaggcacg aacccagttg acataagcct gttcggttcg taaactgtaa
     2821 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     2881 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca gtctatgcct
     2941 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     3001 aacgatgtta cgcagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     3061 aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc aagtcaaatc
     3121 catgcgggct gctcttgatc ttttcggtcg tgagttcggc gacgtagcca cctactccca
     3181 acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat tcatcgcgct
     3241 tgctgccttc ga
//
