LOCUS       sequence-13174-e        3741 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 3741)
  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 3741)
  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: pUT1235-HQ858818
FEATURES             Location/Qualifiers
     source          1..3741
                     /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             1028..1036
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(1298..1397)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1414..1433)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1415..1433)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1438..1454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1438..1454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1556..1575)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1567..2376
                     /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(1634..1653)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2523..3111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3012..3031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3265..3282
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3441..3468)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3560..3646)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3695..3717
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3709..3726
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3710..3726
                     /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 ccccttcacc
      121 atggagggga tggaggaagc aaacagggag gcagtgcaga gctgtcacag ggtgctcacc
      181 ctcctctcca gcccccatag ccagcttgtc cctaacaagg atctcgcggc ggctacggga
      241 gaggctgttg caaagttctg ctccgtggca tccaggctga acaatggcaa tggcctccag
      301 ggccatgccc gggttagaaa gatcaagaag cccttgccta tctttgatag caacctcttt
      361 ttggagagcc ctgcactggc tgtggcgatg gcagccaaga cacctaactc gagtccgatc
      421 actagcctcc agctgtttcc gaggtaccat cagatggagg gctcttcttc aaaggatcct
      481 gttaggatcc ctgcccaatt ccccaaaagg ttgcttctag ataaccctgc ggttgattcg
      541 gatgggccat cacgggggcc accgctccag ctcatccagc cggtgtctgt tgcgccccca
      601 gctggaacgc ctcacccggc attgccttcg gcgcatcttc atttcatcca gcagcatcag
      661 agctaccaga ggtttcagct catgcagcag atgaagatgc agagtgagat gataaagagg
      721 agtggccttg gtgaacaggg tggtagcaat ggtggtggta agggtgttaa tctcaagttt
      781 gacagttcta attgcacggc atcatcctcc cgttcattct tgtcgtctct gagtatggag
      841 gggagcattg cgagtttgga cggtagccgc tccagtcgtc cattccagct agttagtggg
      901 tcacagacgt caagcactcc agagttaggc ctcatgcaga ggaggaggtg cactggtagg
      961 gaggatggta gtggacgatg cacaactggg agccgatgtc actgtgcgaa gaaaaggaag
     1021 ctgaggataa ggaggtctat caaagtccct gcaatcagta ataaggttgc tgacattcca
     1081 gctgatgagt tctcatggag gaagtatgga cagaagccaa taaaaggatc cccacatcct
     1141 aggggttact ataagtgtag cagtgtaaga ggatgcccag ccaggaagca tgttgagagg
     1201 tgcgtcgacg acccctcgat gttgattgtt acctatgaag gtgatcacaa tcacaaccgg
     1261 gtcctagctg cccagccagc caagggtggg cgcgccgacc cagctttctt gtacaaagtt
     1321 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     1381 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     1441 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     1501 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     1561 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     1621 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1681 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1741 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     1801 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     1861 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     1921 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1981 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2041 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2101 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2161 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2221 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2281 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2341 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     2401 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     2461 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2521 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2581 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2641 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2701 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2761 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2821 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2881 acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa
     2941 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3001 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3061 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3121 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3181 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3241 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3301 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3361 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
     3421 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
     3481 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     3541 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     3601 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     3661 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
     3721 gccagtctta agctcgggcc c
//
