LOCUS       sequence-13226-e        3918 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 3918)
  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 3918)
  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: pUT1679-HQ858870
FEATURES             Location/Qualifiers
     source          1..3918
                     /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(1475..1574)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1591..1610)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1592..1610)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1615..1631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1615..1631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1733..1752)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1744..2553
                     /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(1811..1830)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2700..3288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3189..3208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3442..3459
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3618..3645)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3737..3823)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3872..3894
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3886..3903
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3887..3903
                     /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 catggaggat tgcagcagct ggatccatgg ctacgccaac
      181 gctaacgcca ccgccggcaa caacggcttc atgtgcggct acgctgccag ctgcagccca
      241 gtagagtttc agcagcagca acagctggtc ggctcgcaga ttgagcacca cctcaaccag
      301 atcagcatgc agatggggat ggatgacgag tcggcggtgt acgacggcgc ctccatggtg
      361 gacgtcctcc tcatggcttc ctcgtcgccg caccaccacg ccggcgccgg cagcttccag
      421 tactcctcgc cgacgtcctc ctccgcctcc ttccgctccg cctccgtctc gtgcagccct
      481 gagagctcgg cggcggcgac gacgcacttc ctcggaccgc cggcgccgtc cgcggcggcg
      541 gcggggttcc attacccgga ggtctcctcg caggcgccgt tgccactacc cttgccgccc
      601 tacgagccgc agcacggcca atacaccacc gtcctctcgc cgccgccgcc ggcgccagag
      661 ttgccggcga ctactacgcc ggcgaccggc ggcgcgttca ggcggtacgc gcggcacctc
      721 cgcccgagga ggctgcccaa gccgggaggg tgcgggcaga ggatgttcaa gacggccatg
      781 tcggtgctca ccaagatgca cgtggcggcg acgtacaacc gccagtacta ctaccagcag
      841 gcggcagccg ccgccgcgtc ggcgtcggcg gccgaggcgc cgccgtccgg caaccagctg
      901 cagcacatga tctcggagcg gaagcggcgg gagaagctca acgacagctt cctcgccctc
      961 aaggccgtcc tccctcccgg ctctaagaaa gacaagacat cgatactgat cagagcaaga
     1021 gagtacgtaa aatctctcga gtcaaagctg tcagagctgg aggagaagaa ccgggagcta
     1081 gaggcgaggc tagccagccg ccccgccgcc gccgccaaga acgacaaagg cgagacggcg
     1141 gcggcgccag cgccggaagc cggcgatgag acgaagcgaa aggacctagt agagatcgag
     1201 gtgacgacga gcggcggcgg cgccggagcg gcggatgcgg cggcggcggc gggaggagat
     1261 caagagacct gcacgctcaa cgtagacttg cgcggcggcg gaggcggcgg aggcatgagc
     1321 acgacggacg ttgtgctccg gacgctgcag tgcctgagag agcagatcgg cgacggcgcc
     1381 agcctcgtgg cgatgagcac cagcgccggc tccggcggcc gcccgcctcg tgcaaaccta
     1441 acattacagc tcaaggtgaa gggtgggcgc gccgacccag ctttcttgta caaagttggc
     1501 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1561 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1621 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1681 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1741 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1801 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1861 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1921 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1981 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     2041 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     2101 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     2161 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2221 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2281 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2341 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2401 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2461 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2521 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2581 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2641 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2701 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2761 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2821 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2881 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2941 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3001 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3061 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     3121 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3181 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3241 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3301 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3361 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3421 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3481 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3541 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3601 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3661 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3721 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3781 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3841 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3901 agtcttaagc tcgggccc
//
