LOCUS       sequence-79411-e        4491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible entry vector for direct cloning of artificial 
            microRNAs into Arabidopsis thaliana MIR390a precursor .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4491)
  AUTHORS   Carbonell A, Takeda A, Fahlgren N, Johnson SC, Cuperus JT, 
            Carrington JC
  TITLE     New Generation of Artificial MicroRNA and Synthetic Trans-Acting 
            Small Interfering RNA Vectors for Efficient Gene Silencing in 
            Arabidopsis.
  JOURNAL   Plant Physiol. 2014 Mar 19.
  PUBMED    24647477
REFERENCE   2  (bases 1 to 4491)
  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: "Plant 
            Physiol. 2014 Mar 19."
COMMENT     Sequence Label: pENTR-AtMIR390a-B/c
FEATURES             Location/Qualifiers
     source          1..4491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(962..967,968..985,986..992)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1046..1705
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1112..1131)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2047..2352
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2231..2250
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2616..2715)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2732..2751)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2733..2751)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2756..2772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2756..2772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2874..2893)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2885..3694
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2952..2971)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3841..4429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4330..4349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccta taggggggaa aaaaaggtag tcatcagata
      721 tatattttgg taagaaaata tagaaatgaa taatttcacg tttaacgaag aggagatgac
      781 gtgtgttcct tcgaacccga gttttgttcg tctataaata gcaccttctc ttctccttct
      841 tcctcacttc catcttttta gcttcactat ctctctataa tcggttttat ctttctctaa
      901 gtcacaaccc aaaaaaacaa agtagagaag aatctgtaag agaccattag gcaccccagg
      961 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg agccgtcgag
     1021 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     1081 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     1141 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1201 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1261 gttccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1321 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     1381 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1441 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     1501 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1561 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1621 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1681 cgatgagtgg cagggcgggg cgtaaacgcg tggagccggc ttactaaaag ccagataaca
     1741 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     1801 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     1861 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     1921 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     1981 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
     2041 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     2101 gatgtacaga gtgatattat tgacacgccc ggccgacgga tggtgatccc cctggccagt
     2161 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     2221 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtttccgt tatcggggaa
     2281 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     2341 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcacctcg acggtctcac
     2401 attggctctt cttactacaa tgaaaaaggc cgaggcaaaa cgcctaaaat cacttgagaa
     2461 tcaattcttt ttactgtcca tttaagctat cttttataaa cgtgtcttat tttctatctc
     2521 ttttgtttaa actaagaaac tatagtattt tgtctaaaac aaaacatgaa agaacagatt
     2581 agatctcatc tttagtctca agggtgggcg cgccgaccca gctttcttgt acaaagttgg
     2641 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2701 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2761 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2821 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2881 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2941 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     3001 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     3061 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     3121 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     3181 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3241 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3301 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3361 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3421 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3481 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3541 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3601 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3661 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3721 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3781 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3841 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3901 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3961 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4021 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4081 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4141 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4201 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     4261 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4321 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4381 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4441 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
