LOCUS       sequence-13203-e        3396 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 3396)
  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 3396)
  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: pUT1610-HQ858847
FEATURES             Location/Qualifiers
     source          1..3396
                     /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(953..1052)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1069..1088)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1070..1088)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1093..1109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1093..1109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1211..1230)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1222..2031
                     /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(1289..1308)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2178..2766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2667..2686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2920..2937
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3096..3123)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3215..3301)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3350..3372
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3364..3381
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3365..3381
                     /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 catggacccc gcccccacgc tcgccgccga gctctggcga
      181 acgccgtacc tcggtggcgg tggtggtgga ggaggaggag ggcgtgggtt ggaggcggcg
      241 gcgtccgggg tcacggagca gagcaatggg agccgcgggg gtggcggagg aggaggcgca
      301 gggcggagga ggcagcggga ggcccccgcg ctcgaggacg actcgtccag gatcgtctcc
      361 gcttccggag gaggaggcgg cggtcaggat ttgactgatt cagaagcaaa gcgtttcaag
      421 gcaagtaaat caagtggaga taatagcagt ttaagaacag aggctgaaac tgattcaaga
      481 aatgctagca agtctggtga tcaaaatccg ccaccaccag aaccaccaaa gcaagactac
      541 atccatgtga gggcaagaag gggtcaagct actgacagcc atagccttgc cgaaagggca
      601 cgtcgtgaga aaataagtga gcggatgaaa attctccagg atcttgtccc tggatgcaac
      661 aaggttattg ggaaagcatc agtacttgat gagataatta attatatcca ggctttgcaa
      721 cggcaagttg agtttttgtc catgaagctt gaggctgtta atgctcatgt gaataatgga
      781 atcgaagcat ttccacccaa agattttggt gctcaggtat acaacacggc tcctggattg
      841 acgtttgatc cacaaacacc aagagaatac gcccagggat caacaccatc ggaatggctc
      901 cacatgcaga ttggtggcac ctatgaaaga gtgacaaagg gtgggcgcgc cgacccagct
      961 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1021 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1081 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1141 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1201 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1261 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1321 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1381 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1441 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1501 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1561 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1621 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1681 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1741 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     1801 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     1861 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     1921 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     1981 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2041 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2101 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2161 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2221 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2281 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2341 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2401 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2461 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2521 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     2581 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2641 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2701 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2761 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2821 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2881 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2941 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3001 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3061 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3121 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3181 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3241 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3301 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     3361 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
