LOCUS       sequence-233129-        4020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone containing AtMIR390a backbone (split by ccdB gene). Used
            to make artificial microRNA construct. For use in plants and 
            compatible with the MultiSite Gateway system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4020)
  AUTHORS   Wang X, Ye L, Lyu M, Ursache R, Loytynoja A, Mahonen AP
  TITLE     An inducible genome editing system for plants.
  JOURNAL   Nat Plants. 2020 Jun 29. pii: 10.1038/s41477-020-0695-2. doi: 
            10.1038/s41477-020-0695-2.
  PUBMED    32601420
REFERENCE   2  (bases 1 to 4020)
  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: "Nat Plants.
            2020 Jun 29. pii: 10.1038/s41477-020-0695-2. doi: 
            10.1038/s41477-020-0695-2."
COMMENT     Sequence Label: p221z-AtMIR390a
FEATURES             Location/Qualifiers
     source          1..4020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(125..152)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(244..330)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     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"
     protein_bind    426..525
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(819..824,825..842,843..849)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             903..1562
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(969..988)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1904..2209
                     /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     2088..2107
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2473..2572)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2589..2608)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2590..2608)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2613..2629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2613..2629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2697..3071)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(3090..3137)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      3227..3815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3716..3735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3969..3986
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
      121 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
      181 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
      241 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      301 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      361 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      421 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
      481 agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccgcg gccgccccct
      541 tcacctatag gggggaaaaa aaggtagtca tcagatatat attttggtaa gaaaatatag
      601 aaatgaataa tttcacgttt aacgaagagg agatgacgtg tgttccttcg aacccgagtt
      661 ttgttcgtct ataaatagca ccttctcttc tccttcttcc tcacttccat ctttttagct
      721 tcactatctc tctataatcg gttttatctt tctctaagtc acaacccaaa aaaacaaagt
      781 agagaagaat ctgtaagaga ccattaggca ccccaggctt tacactttat gcttccggct
      841 cgtataatgt gtggattttg agttaggagc cgtcgagatt ttcaggagct aaggaagcta
      901 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
      961 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1021 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     1081 ttcacattct tgcccgcctg atgaatgctc atccggagtt ccgtatggca atgaaagacg
     1141 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     1201 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     1261 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     1321 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     1381 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     1441 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     1501 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     1561 aaacgcgtgg agccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     1621 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc
     1681 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     1741 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     1801 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     1861 gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta
     1921 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     1981 cacgcccggc cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     2041 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     2101 cgatatggcc agtgtgccgg tttccgttat cggggaagaa gtggctgatc tcagccaccg
     2161 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     2221 ccttatacac agccagtctg cacctcgacg gtctcacatt ggctcttctt actacaatga
     2281 aaaaggccga ggcaaaacgc ctaaaatcac ttgagaatca attcttttta ctgtccattt
     2341 aagctatctt ttataaacgt gtcttatttt ctatctcttt tgtttaaact aagaaactat
     2401 agtattttgt ctaaaacaaa acatgaaaga acagattaga tctcatcttt agtctcaagg
     2461 gtgggcgcgc cgacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
     2521 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     2581 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     2641 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgatcag
     2701 tcctgctcct cggccacgaa gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc
     2761 cgcccccacg gctgctcgcc gatctcggtc atggccggcc cggaggcgtc ccggaagttc
     2821 gtggacacga cctccgacca ctcggcgtac agctcgtcca ggccgcgcac ccacacccag
     2881 gccagggtgt tgtccggcac cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg
     2941 tcccggacca caccggcgaa gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg
     3001 gtccagaact cgaccgctcc ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc
     3061 aacttggcca tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc
     3121 gatgattaat tgtcaacacg tgctgatcat gaccaaaatc ccttaacgtg agttacgcgt
     3181 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3241 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3301 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3361 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3421 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3481 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3541 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3601 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3661 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3721 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3781 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3841 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3901 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3961 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
//
