LOCUS       sequence-13209-e        3618 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 3618)
  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 3618)
  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: pUT1616-HQ858853
FEATURES             Location/Qualifiers
     source          1..3618
                     /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)"
     regulatory      305..314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1175..1274)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1291..1310)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1292..1310)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1315..1331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1315..1331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1433..1452)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1444..2253
                     /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(1511..1530)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2400..2988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2889..2908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3142..3159
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3318..3345)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3437..3523)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3572..3594
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3586..3603
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3587..3603
                     /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 catggagagc ttgtgtgggc agcttgagca tgaacatgag
      181 cagcagctgc accaatcctc ctcatcttca cttctcccct actgcttcct ttctgctcca
      241 gagcctcttc ctgtcccagg cttccctgcc ggctgcagca accttgaaga gaaggcagca
      301 gcagccatgg cggcctatga gtatgagagc agcagctgca gctcccttga ccccacctca
      361 atgccaatgg tgtactcacc aatagtgctg cagccacagg agtgccctct ttcctttgtc
      421 tttgacaatg ccgctgccgc tgccggcgac aacaaatggg tgcctggcat tcaaggaagt
      481 tgcccttgtt cattgggctc aactcaagat atggatgcaa gttggggcaa gagtagaaag
      541 cacaagagga gcaatgtggg cttgaaggga cttgaggaga aaaaggcaag aagggtggta
      601 ctacatcaac atgatgatga tgtgaagaag aaggccaagg aggcagcagg tggggagcca
      661 ccagctgggt acattcatgt gagggcaagg agaggccaag caacagacag ccacagcctt
      721 gctgagaggg tgagaaggga gaagatcagt gagaggatga agatgctgca aagcctggtt
      781 cctggttgtg acaaggtcac agggaaagcc ctcatgctag atgagatcat cagctatgtg
      841 caatctctgc agaaccaagt tgagtttctg tccatgaagc ttgcctccct tagccctttg
      901 atgtatgagt ttggaccggg cattgatatg cacccagatg ttcttcgaca gcttgcaaag
      961 atgccacatg agatggtgca gtgcatgggc caaatgggga gtacaggcat ttcactccaa
     1021 ggactaggag gaggacccac aggctttgct caggatggca gcagccacat taacatggtg
     1081 gtgatgcaag ttggagagca aggccagcag cagggctctc ttcaccaggt cgagatgagc
     1141 agccactgct tctttcataa gggtgggcgc gccgacccag ctttcttgta caaagttggc
     1201 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1261 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1321 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1381 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1441 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1501 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1561 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1621 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1681 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1741 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1801 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1861 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1921 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1981 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2041 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2101 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2161 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2221 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2281 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2341 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2521 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2761 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3181 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3241 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3301 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3361 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3421 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3481 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3541 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3601 agtcttaagc tcgggccc
//
