LOCUS       sequence-100047-        4122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible entry vector for direct cloning of artificial 
            microRNAs into rice MIR390 precursor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4122)
  AUTHORS   Carbonell A, Fahlgren N, Mitchell S, Cox KL Jr, Mockler TC, 
            Carrington JC
  TITLE     Highly Specific Gene Silencing in a Monocot Species by Artificial 
            MicroRNAs Derived From Chimeric MIRNA Precursors.
  JOURNAL   Plant J. 2015 Mar 24. doi: 10.1111/tpj.12835.
  PUBMED    25809382
REFERENCE   2  (bases 1 to 4122)
  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 J. 
            2015 Mar 24. doi: 10.1111/tpj.12835."
COMMENT     Sequence Label: pENTR-OsMIR390-B/c
FEATURES             Location/Qualifiers
     source          1..4122
                     /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(753..758,759..776,777..783)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             837..1496
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(903..922)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1838..2143
                     /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     2022..2041
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2247..2346)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2363..2382)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2364..2382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2387..2403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2387..2403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2505..2524)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2516..3325
                     /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(2583..2602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3472..4060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3961..3980
                     /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 ccttcaccga gctcgagatg ttttgaggaa gggtatggaa
      721 caatccttga gagaccatta ggcaccccag gctttacact ttatgcttcc ggctcgtata
      781 atgtgtggat tttgagttag gagccgtcga gattttcagg agctaaggaa gctaaaatgg
      841 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
      901 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
      961 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     1021 ttcttgcccg cctgatgaat gctcatccgg agttccgtat ggcaatgaaa gacggtgagc
     1081 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     1141 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     1201 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     1261 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     1321 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     1381 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     1441 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaaacgc
     1501 gtggagccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     1561 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa
     1621 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     1681 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     1741 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     1801 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     1861 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     1921 cggccgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     1981 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     2041 ggccagtgtg ccggtttccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     2101 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     2161 acacagccag tctgcacctc gacggtctca catggtttgt tcttaccaca cgaccaatta
     2221 aatcgagctc aagggtgggc gcgccgaccc agctttcttg tacaaagttg gcattataag
     2281 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2341 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2401 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2461 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2521 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2581 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2641 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2701 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2761 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2821 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2881 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2941 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3001 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3061 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3121 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3181 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3241 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3301 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3361 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3421 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3601 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3841 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     3901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4081 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
