LOCUS       sequence-79409-e        4989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible entry vector for direct cloning of synthetic 
            trans-acting siRNAs into Arabidopsis thaliana TAS1c precursor .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  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 4989)
  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-AtTAS1c-B/c
FEATURES             Location/Qualifiers
     source          1..4989
                     /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(1132..1137,1138..1155,1156..1162)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1216..1875
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1282..1301)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2217..2522
                     /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     2401..2420
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3114..3213)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3230..3249)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3231..3249)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3254..3270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3254..3270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3372..3391)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3383..4192
                     /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(3450..3469)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4339..4927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4828..4847
                     /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 ccttcaccaa acctaaacct aaacggctaa gcccgacgtc
      721 aaataccaaa aagagaaaaa caagagcgcc gtcaagctct gcaaatacga tctgtaagtc
      781 catcttaaca caaaagtgag atgggttctt agatcatgtt ccgccgttag atcgagtcat
      841 ggtcttgtct catagaaagg tactttcgtt tacttctttt gagtatcgag tagagcgtcg
      901 tctatagtta gtttgagatt gcgtttgtca gaagttaggt tcaatgtccc ggtccaattt
      961 tcaccagcca tgtgtcagtt tcgttccttc ccgtcctctt ctttgatttc gttgggttac
     1021 ggatgttttc gagatgaaac agcattgttt tgttgtgatt tttctctaca agcgaataga
     1081 ccatttatcg gtggatctta gaaaattaag agaccattag gcaccccagg ctttacactt
     1141 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg agccgtcgag attttcagga
     1201 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     1261 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     1321 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1381 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga gttccgtatg
     1441 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     1501 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     1561 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     1621 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     1681 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     1741 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     1801 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     1861 cagggcgggg cgtaaacgcg tggagccggc ttactaaaag ccagataaca gtatgcgtat
     1921 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     1981 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     2041 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     2101 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     2161 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
     2221 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     2281 gtgatattat tgacacgccc ggccgacgga tggtgatccc cctggccagt gcacgtctgc
     2341 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     2401 gcatgatgac caccgatatg gccagtgtgc cggtttccgt tatcggggaa gaagtggctg
     2461 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     2521 aaatgtcagg ctcccttata cacagccagt ctgcacctcg acggtctcag aactagaaaa
     2581 gacattggac atattccagg atatgcaaaa gaaaacaatg aatattgttt tgaatgtgtt
     2641 caagtaaatg agattttcaa gtcgtctaaa gaacagttgc taatacagtt acttatttca
     2701 ataaataatt ggttctaata atacaaaaca tattcgagga tatgcagaaa aaaagatgtt
     2761 tgttattttg aaaagcttga gtagtttctc tccgaggtgt agcgaagaag catcatctac
     2821 tttgtaatgt aattttcttt atgttttcac tttgtaattt tatttgtgtt aatgtaccat
     2881 ggccgatatc ggttttattg aaagaaaatt tatgttactt ctgttttggc tttgcaatca
     2941 gttatgctag ttttcttata ccctttcgta agcttcctaa ggaatcgttc attgatttcc
     3001 actgcttcat tgtatattaa aactttacaa ctgtatcgac catcatataa ttctgggtca
     3061 agagatgaaa atagaacacc acatcgtaaa gtgaaataag ggtgggcgcg ccgacccagc
     3121 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     3181 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     3241 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     3301 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     3361 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     3421 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     3481 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     3541 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     3601 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3661 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3721 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3781 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3841 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3901 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3961 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     4021 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     4081 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     4141 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     4201 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     4261 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     4321 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4381 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4441 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4501 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4561 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4621 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4681 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     4741 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4801 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4861 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4921 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4981 tcacatgtt
//
