LOCUS       sequence-13227-e        3498 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 3498)
  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 3498)
  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: pUT1684-HQ858871
FEATURES             Location/Qualifiers
     source          1..3498
                     /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(1055..1154)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1171..1190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1172..1190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1195..1211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1195..1211)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1313..1332)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1324..2133
                     /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(1391..1410)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2280..2868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2769..2788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3022..3039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3198..3225)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3317..3403)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3452..3474
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3466..3483
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3467..3483
                     /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 catgcaaatg gagtcgtact acggcgcgtt ccacgccgac
      181 gaggccgcgt tcttcttccc ccaccacgtg ccggcgtcgc cggagctgcc gttcggcctc
      241 attgcgtcgc cggagccgga gccggagccg gagcaggcgg cggcggaggc gaggcagagc
      301 gcgttccagg agtacggcgg cgcggtgcac gccggagcgc cggcggcggc gggggctgtc
      361 accaccggtg ggacgaacat ccaccggagg gtgatggacg tgctgggcag gatgggcggc
      421 ggcggcggcg ggggggagaa gggggagggt gaggagatgg aggaggagga ggaggtgccg
      481 cagcggcggc ggcgcgggca gggcgccgac gtggagagca gccgcggttt ccgccacatg
      541 atgcgcgagc gccagcgccg cgagaagctc agccagagct acgccgacct ctacgccatg
      601 gtctcctctc gctccgaggg ggacaagaac tcgatcgtgc agtcggcggc gatctacatc
      661 cacgagctga aggtcgcgag ggaccagctg cagaggagga acgaggagct gaaggcccag
      721 atcatgggcc acgacgagca gcagccgtgc gtcacggtgc agttcgaggt cgacgagccg
      781 tcgtcgtcga tcgactccat gatcgccgcg ctccggcggc tcaagagcat gagcgtcaag
      841 gcgcgcggga tccggtcgag catgtccggg aacaggctgt ggacggagat gaacgtcgag
      901 accacgattg cggcttgtga agtggaaaag gcagtggaag aggctctcaa ggaagtagag
      961 aggaaccagc ctgacagcga tgccccattt cctggaagca aaggctggac acagacatct
     1021 catgtgcaaa atgtgtttaa gggtgggcgc gccgacccag ctttcttgta caaagttggc
     1081 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1141 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1201 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1261 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1321 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1381 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1441 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1501 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1561 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1621 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1681 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1741 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1801 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1861 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1921 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1981 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2041 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2101 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2161 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2221 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2281 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2341 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2401 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2461 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2521 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2581 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2641 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2701 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2761 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2821 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2881 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2941 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3001 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3061 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3121 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3181 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3241 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3301 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3361 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3421 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3481 agtcttaagc tcgggccc
//
