LOCUS       sequence-13231-e        3183 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 3183)
  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 3183)
  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: pUT1752-HQ858875
FEATURES             Location/Qualifiers
     source          1..3183
                     /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      140..149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(740..839)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(856..875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(857..875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(880..896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(880..896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(998..1017)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1009..1818
                     /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(1076..1095)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1965..2553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2454..2473
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2707..2724
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2883..2910)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3002..3088)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3137..3159
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3151..3168
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3152..3168
                     /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 catggcggct tccgtaggac tgaaccctga agctttcttc
      181 ttcagcaact cctactccta ctcctcatcc cctttcatgg ccagctacac gccggagttc
      241 tcggccgccg ctatcgacgc caacttattc tccggcgagc tcgatttcga ctgctctctc
      301 ccagctccgg cccaggagta cccggaaaat gaaaacacta tgatgaggta cgagagcgag
      361 gagaagatga gggcgagggt gaacgggagg atcgggttca ggacgaggtc ggaggtggag
      421 attcttgatg atggcttcaa gtggaggaag tacggcaaga aagctgtcaa gaacagccca
      481 aatccaagaa actactaccg gtgctcgacg gaggggtgca acgtgaagaa gcgagtggag
      541 agagaccggg aggaccaccg ctacgtcatc accacctacg acggcgtcca caaccacgcg
      601 agccccgccg ccgccgccgc ggcgctgcag tacgccgccg ccgccggcga ctactacagc
      661 ccgccgctca gcagcgccgt ctcgccgccg gccgcctatt cggcaggcgg ctcgctgctc
      721 ttcaagggtg ggcgcgccga cccagctttc ttgtacaaag ttggcattat aagaaagcat
      781 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
      841 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
      901 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
      961 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     1021 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1081 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     1141 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1201 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1261 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     1321 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1381 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1441 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1501 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     1561 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     1621 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     1681 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     1741 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     1801 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     1861 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     1921 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1981 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2041 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2101 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2161 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2221 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2281 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2341 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2401 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2461 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2521 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2581 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2641 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2701 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     2761 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     2821 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     2881 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     2941 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     3001 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     3061 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     3121 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     3181 ccc
//
