LOCUS       sequence-13229-e        3336 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 3336)
  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 3336)
  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: pUT1750-HQ858873
FEATURES             Location/Qualifiers
     source          1..3336
                     /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(893..992)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1009..1028)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1010..1028)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1033..1049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1033..1049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1151..1170)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1162..1971
                     /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(1229..1248)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2118..2706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2607..2626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2860..2877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3036..3063)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3155..3241)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3290..3312
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3304..3321
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3305..3321
                     /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 catgtacatg gcggcggcgg ctgcgggagc aagcacaccc
      181 ttcaattttt gccgccacgg ctcgcacgcc gagtacgacg ccgtcttcag cggcagctgg
      241 atggcgcgcc gcccctccgc ggcgccgcac ggtggcggcg cctccggctc cggctccggc
      301 agcggctacg gcgccgcctc gtacgttgct cccaccttcg gcgcggcgtt taggcagcag
      361 catctcgacc tcctcgacta cctgtccgac gatcagggcg ttccggcgcc accaccagcg
      421 gctgtgccca gcgcgtcgta cgtgacgccg gcgccggcga tggcccccgc tgaaccggtc
      481 gtgccggatg ctgttgccgc cgccggcggc tatccgagga gcgtggctgc ggcggcggcg
      541 gccgtggccg gcgagggaag agatcggacg acgacggaca agatcgcatt ccggacgagg
      601 tcggacgatg agatactcga cgacgggtat aagtggagga agtacgggaa aaagtccgtt
      661 aagaacagcc ctaatccgag gaactactac cggtgctcga cggaggggtg caacgtgaag
      721 aagagggtgg agagagacaa gaacgacccg cggtacgtgg tgacgatgta cgaggggatc
      781 cacaaccacg tgtgccccgg cactgtctac tacgccgcgc aggacgccgc ctccggccgc
      841 tttttcgtcg ccgggatcag tcacccagac cttaataagg gtgggcgcgc cgacccagct
      901 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
      961 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1021 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1081 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1141 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1201 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1261 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1321 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1381 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1441 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1501 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1561 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1621 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1681 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     1741 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     1801 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     1861 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     1921 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     1981 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2041 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2101 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2161 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2221 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2281 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2341 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2401 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2461 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     2521 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2581 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2641 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2701 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2761 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2821 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2881 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2941 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3001 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3061 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3121 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3181 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3241 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     3301 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
