LOCUS       sequence-13225-e        3816 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 3816)
  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 3816)
  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: pUT1678-HQ858869
FEATURES             Location/Qualifiers
     source          1..3816
                     /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(1373..1472)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1489..1508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1490..1508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1513..1529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1513..1529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1631..1650)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1642..2451
                     /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(1709..1728)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2598..3186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3087..3106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3340..3357
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3516..3543)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3635..3721)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3770..3792
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3784..3801
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3785..3801
                     /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 catggacggc ggcagggtcg ggggcgacta catttccagc
      181 ctcctcagct cctccccccg gctcgacttc ggcgtgccgg tgctggacgc catcgtggcg
      241 ccgggaggcg gcggcggcgg cggcgactgc ggcctcgaca agctctgcgg cgacccggtg
      301 ttcgcggagc gcgccgcgcg gctgtccagc ttcaacaacg gcggcggcgg tgttgggcag
      361 cggtacggcg gcgcgggggc ggggctcttc gggatgcctc ctccggctcc cggcgacttc
      421 gccggaggtg gctccaggga ggcgtcctcc gtgtccgacc ccgcctcctc ggcgatgaag
      481 gacgccgccg ccaatgccaa gaagaggaag tccacggcgg cggcggccgc cgccgccaag
      541 ggcaaaggca aggagcctcc ggttggggaa gagaaggaat cagatggcaa gaggtgcaag
      601 acgggcaatg gcgagaagga gagctcggtg aagccgaagg cggagcaggc cgggagcgac
      661 agctccgtcg aggacggcgg cggtggcggg cagaagcaag ggaaggggaa gaatgccaag
      721 ccggtggagc cacccaagga ctatgtccat gtccgggcga ggcgaggcca agccactgac
      781 agccacagcc tcgctgagag ggtcagaaga gagaggatca gccagaggat gaaggttctc
      841 caggacctgg ttccaggatg caacaaggta attggcaagg cactcatgct tgatgagatc
      901 ataaactacg tgcaatcgct tcaacggcaa gttgagttcc tgtccatgaa gctagcaact
      961 gtgaatccac ttgatttcag taacctgcct actctgctgc agaaagatat gctccaagca
     1021 tgcggccctt cagcgagctc ggttttctca ttagaaagct ccaattctgc ttttcggttt
     1081 gccgaacaag gagatgtttt ccaacagttt gctcaaaaca gcatggaaag ccagtgcacc
     1141 ctgaatcagt tggacctggc tctatctcag gctacaaatg ctgcacagta tgcttttcaa
     1201 gatggaacag ccggcgcaaa cttgcagcaa aggaacttct gggaggatga tctccaaagt
     1261 gttttccata ttgagaatgg acagagccag gagaatgggg tttcagcacc aaacttccat
     1321 ggtcagcagc aagcaggcca catgaaaatg gagttcaagg gtgggcgcgc cgacccagct
     1381 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1441 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1501 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1561 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1621 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1681 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1741 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1801 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1861 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1921 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1981 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2041 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2101 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2161 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     2221 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2281 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2341 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2401 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2461 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2521 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     3001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3301 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3361 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3421 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3481 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3541 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3601 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3661 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3721 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     3781 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
