LOCUS       sequence-13232-e        3564 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 3564)
  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 3564)
  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: pUT1754-HQ858876
FEATURES             Location/Qualifiers
     source          1..3564
                     /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)"
     protein_bind    complement(1121..1220)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1237..1256)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1238..1256)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1261..1277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1261..1277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1379..1398)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1390..2199
                     /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(1457..1476)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2346..2934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2835..2854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3088..3105
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3264..3291)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3383..3469)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3518..3540
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3532..3549
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3533..3549
                     /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 catggtgaag gaggaggaga tcatcacaga ggcagcagga
      181 gcaagaggct acatggagat gcttgggctt ggagaggagg ccgccgacta cctgatgtgc
      241 ctgtctcctt cttcctacct gtcttctcct gctgcttcca ccaccacagc tgtggcttct
      301 cctacctgtg cctcgtactt ggctcctcac ccttaccacc acctgctaag cttcagtggt
      361 caggatcagt accacggtga tgacgtcttt gggttacagt attacggcgg cgatcaagta
      421 attccggcgg tggttccgca gaagtccagc ccgaccaccg agtgctcctc ctccgtctcc
      481 tccatgtcgt cctcgccgac ggccaccgcc atttccagct ctaagtcacc ggccttcaag
      541 aagaagggat cgagaggttg tgatcagagg aaggctactg ctcctgctgc tgccacaacc
      601 acaaacaaga gacccagggt gaggagggag aggctcggag agaggatcat agcgctgcag
      661 cagctggttt ctccgttcgg gaagtctgat acggcctccg ttcttcacga ggcgttggga
      721 tacatacgct tcctgcacga ccaggttcag gtcctgagct cgccgtacct gcagcgcctg
      781 cctccctctg ctcgcgtccc ggagcaggag agaggcacgc cggcggcgga ggagcaaccg
      841 ccgccactgc cgcattcgct gcggccgagc gacctgagga gccgtgggct gtgcctggtg
      901 ccgatctcgt gcaccgagca tgtcgccggc gccggcgccg gcaccggcca cggcaacggc
      961 gccgacctat ggtcggtggc ggcgggcatg gcgaaggcga cggcgaccgt gacggccgcc
     1021 gtggagagga gcaaggaggc agccgccgcc gccgccgcca ctgccgcatt gctgcgtgct
     1081 gatcgtccag ggcagcagct ggccaagggt gggcgcgccg acccagcttt cttgtacaaa
     1141 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1201 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1261 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1321 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1381 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1441 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1501 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1561 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1621 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1681 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1741 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1801 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1861 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1921 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1981 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2041 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2101 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2161 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2221 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2281 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2341 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2401 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2461 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2521 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2581 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2641 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2701 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     2761 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2821 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2881 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2941 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3001 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3061 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3121 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3181 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3241 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     3301 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     3361 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     3421 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     3481 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     3541 acggccagtc ttaagctcgg gccc
//
