LOCUS       sequence-13223-e        3483 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 3483)
  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 3483)
  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: pUT1676-HQ858867
FEATURES             Location/Qualifiers
     source          1..3483
                     /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(1040..1139)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1156..1175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1157..1175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1180..1196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1180..1196)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1298..1317)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1309..2118
                     /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(1376..1395)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2265..2853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2754..2773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3007..3024
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3183..3210)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3302..3388)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3437..3459
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3451..3468
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3452..3468
                     /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 catggccggg cagcagccgc agcagcaggg cccaccggag
      181 gacgactttt tcgaccagtt cttctccctg accagctcct tccctggcgc cgcgccgggc
      241 ggccgcgccg ccggtgacca gcccttctcc ctcgcgctca gcctcgacgc cgccgcggcg
      301 gccgaggcgt ccgggagcgg gaagaggctc ggggtcggcg atgacgccga gggtggcggc
      361 agcaaggcgg atcgggagac cgtgcagctc accggactct tcccgccggt gttcggcggc
      421 ggcggcgtgc agccgccgaa cctccgcccc accccgccta cccaggtgtt ccacccgcag
      481 cagtcgaagc agggcggagc agccgtcggg ccgcagccgc cggcgccgag gccgaaggtg
      541 cgagcgcggc gtgggcaggc gaccgacccc cacagcatcg cggagaggct aagaagagag
      601 agaatagcag aaaggatgag ggccctacag gaattggtcc ccaatacaaa caagacagat
      661 agggcagcta tgctagatga gatccttgat tatgtgaaat tcctgaggct gcaagtaaag
      721 gttttaagca ttagcaggct gggtggcgcg ggtgctgttg cacagctggt tgctgatatt
      781 ccactttcag ttaaggggga agcaagcgat agtgggggca accaacagat ttgggaaaag
      841 tggtcaacgg atggcacaga aagacaggta gcgaagctga tggaagaaga catcggggca
      901 gcgatgcaat ttctccaatc caaagcactc tgcatgatgc caatctcgct cgccatggca
      961 atctatgaca cacaacaaac acaggatgga caaccagtga agcacgaacc caacactcct
     1021 tccaagggtg ggcgcgccga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     1081 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1141 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     1201 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     1261 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     1321 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1381 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     1441 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1501 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1561 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     1621 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1681 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1741 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1801 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     1861 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     1921 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     1981 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2041 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2101 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     2161 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     2221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2401 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2641 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2941 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3001 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3061 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3121 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     3181 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     3241 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     3301 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     3361 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     3421 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     3481 ccc
//
