LOCUS       sequence-13221-e        4125 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 4125)
  AUTHORS   Yilmaz A, Nishiyama MY Jr, Fuentes BG, Souza GM, Janies D, Gray J, 
            Grotewold E
  TITLE     GRASSIUS: a platform for comparative regulatory genomics across the 
            grasses.
  JOURNAL   Plant Physiol. 2009 Jan;149(1):171-80. doi: 10.1104/pp.108.128579. 
            Epub 2008 Nov 5.
  PUBMED    18987217
REFERENCE   2  (bases 1 to 4125)
  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"; doi: 
            "10.1104/pp.108.128579"; journalName: "Plant Physiol"; date: 
            "2009-01"; volume: "149"; issue: "1"; pages: "171-80"
COMMENT     Sequence Label: pUT1674-HQ858865
FEATURES             Location/Qualifiers
     source          1..4125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             112..132
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             759..770
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(1682..1781)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1798..1817)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1799..1817)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1822..1838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1822..1838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1940..1959)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1951..2760
                     /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(2018..2037)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2907..3495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3396..3415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3649..3666
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3825..3852)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3944..4030)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4079..4101
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4093..4110
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4094..4110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc cttgtttaac
      121 tttaagaagg agcccttcac catgggactg caagggaata aggcgacgca cgacttcctc
      181 tcgctgtacg ccgccgccgc caccgccacg gactactcgc cgctcccgcg ccacccggac
      241 tccaagccct cggcgccggc gccgccgccc gctcaagatt tcttcctcaa gacacacgac
      301 ttcctgccgc aggtagagaa gagagccgac cagacaccac cgccgccgcc gccgccgccg
      361 ccccacgccg cggtcaccgc cgagaagcag ctgctccacc agcacgcgtt ggccgccgcc
      421 ggcgcgttca ccatcaacca cgccgtggcc gcggccgcca ccgccgtcaa gcaggagccg
      481 cccttcgcgc cgtggtgcca gcccgtcgcc gccgtagatc cccgaggtca ccagtggtcg
      541 ctccccttcg ccgcgcgcgc cgtcgccgtc gcctcctcca ggccgcagca gcagcagcag
      601 cagccaccgc cgccggagag gaagggtggc ggtggattca tggacgccgg ctccagatcc
      661 agtggcggcg cgggcttcga cgacgacgac ggccatgccg cgcgccgcga ggtctcctcc
      721 tcgctgaaag agctcacggt cagggtggaa gggaaaggtg ggagctgcag cggcagcgct
      781 ggcacggatc agatgccgaa cacgccgcgc tcgaagcact ccgccaccga gcagcgcagg
      841 cgcagcaaaa tcaacgacag atttcaactt cttagagact tattgcctca taatgatcaa
      901 aagagagaca aagcatcatt tctcttagag gttattgaat acatacggtt tttacaagag
      961 aaagtacaaa agtatgaggt ttcatacccg gagtggaacc aagaaaatgc aaaggttgtg
     1021 ccatggacaa acatctattt ccgttcttcc tggaaaaatg cgcagaacaa aggccaagtt
     1081 cctgcagatc attcacctga ccctccagaa cttctgaaaa atggttcccc ctatatgttc
     1141 ccttttacag ggaactctga taataacaat gcagttgaaa ctgcagctgc ttcaggtgca
     1201 caggatcagg cagaaactga tccaatgagt cgtgtgtctt acagatcagt ggacactccc
     1261 agtcccaata atgttgcaga caaagttaca tctcagccac acgctcaact ggtgagacca
     1321 tcacctgcag agaatcacac agtaaactgc gacaagttaa ataattctga ccttgccatc
     1381 gatgagggaa caatcagtct atcaagccaa tattcccaag agttgcttaa caagttgaat
     1441 catgccctgg aaaactcggg tatagatttg tcccaagcca gcatctccgt gcaaataaat
     1501 cttggcaaga gggctatgaa gagatctact cctgctgcaa cctccacttc taaggagctc
     1561 actgatccgg catctaatag tcaagcaatg ggccgtcagc ttaggttggg tgatggcgct
     1621 gaagaacatc gacaagcctc aaagcgacac aaatcagata acagcaaggg tgggcgcgcc
     1681 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     1741 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     1801 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     1861 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1921 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1981 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     2041 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     2101 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     2161 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     2221 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     2281 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     2341 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     2401 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     2461 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     2521 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     2581 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2641 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2701 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     2761 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     2821 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2881 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2941 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3001 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3061 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3121 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3181 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3241 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
     3301 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3361 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3421 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3481 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3541 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3601 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3661 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3721 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3781 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3841 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3901 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3961 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     4021 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     4081 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
