LOCUS       sequence-13234-e        3744 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 3744)
  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 3744)
  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: pUT1858-HQ858878
FEATURES             Location/Qualifiers
     source          1..3744
                     /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             277..300
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     protein_bind    complement(1301..1400)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1417..1436)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1418..1436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1441..1457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1441..1457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1559..1578)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1570..2379
                     /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(1637..1656)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2526..3114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3015..3034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3268..3285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3444..3471)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3563..3649)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3698..3720
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3712..3729
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3713..3729
                     /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 catggacgag gcggaggcgg cggcggcggc ggcgaagatg
      181 gacgagctgg cgggaggagg aggaggaggc ggcggcgact ggtcgtacct cgcggcggac
      241 gcgctggcgg cggcgtcgtt cacggcgttc ccgttccacc accaccacca ccaccaccac
      301 cgcgacgtgc tctcggcgtc gaccccgtcg tcgctgctcc tcaacatgga cgcggccacc
      361 gccgcggcca tgttcgactt ccaggcggcc ttcccgtcgt cctccgtccc gccgccgccg
      421 cccacgacga cggcggcgct cccgccgttc cacgacttcg cctcctccaa cccgttcgac
      481 gacgcgccgc ccccgttcct cgcgccgccg gggcagaagc tcgggttcct ggggcccccc
      541 ggcggcgcgt tcggcggcgg gatggggtgg gacgacgacg acgagatcga gcagagcgtg
      601 gacgcctcgt ccatgggtgt ctcggcctcg ctggagaatg ccgcgcccgt ggcggccgga
      661 ggaggcggcg gcggtggcgg cggcggtggg aggggcaaga agaaggggat gccggccaag
      721 aacctcatgg cggagcggcg gcgcaggaag aagctcaacg accgcctcta catgctgcgc
      781 tccgtggtgc ccaagatcag caagatggac agagcttcta tccttggtga tgcaattgag
      841 tacttgaagg agctcctgca gaggattaac gatcttcaca atgaacttga gtctgctcct
      901 agctcttcgc ttactggacc atcatcggct agcttccacc catcaacacc cacattgcag
      961 acgtttcctg gccgcgttaa ggaggaactt tgcccaacct catttccaag ccctagtggt
     1021 caacaagcca cggtcgaagt taggatgagg gaaggtcatg cagtcaatat ccacatgttc
     1081 tgcgcacgca ggcccggcat tctgatgtcc acattgaggg ctctcgacag ccttggcctc
     1141 gggatcgagc aggcggtcat cagctgtttc aatgggtttg caatggatgt tttccgcgcc
     1201 gagcaatgca gggatggtcc tggactcggg cctgaagaaa ttaagaccgt gctcctgcac
     1261 tctgctggcc ttcagaacgc aatgaagggt gggcgcgccg acccagcttt cttgtacaaa
     1321 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1381 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1441 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1501 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1561 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1621 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1681 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1741 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1801 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1861 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1921 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1981 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2041 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2101 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2161 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2221 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2281 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2341 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2401 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2461 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2641 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2881 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     2941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3181 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3241 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3301 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3361 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3421 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     3481 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     3541 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     3601 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     3661 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     3721 acggccagtc ttaagctcgg gccc
//
