LOCUS       sequence-13205-e        3663 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 3663)
  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 3663)
  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: pUT1612-HQ858849
FEATURES             Location/Qualifiers
     source          1..3663
                     /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      353..362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      549..558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1220..1319)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1336..1355)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1337..1355)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1360..1376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1360..1376)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1478..1497)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1489..2298
                     /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(1556..1575)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2445..3033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2934..2953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3187..3204
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3363..3390)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3482..3568)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3617..3639
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3631..3648
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3632..3648
                     /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 catggagctt gacgaggagt ccttcttgga tgagctcatg
      181 tcgctgcggc gtgatggatc ggcgccgtgg caggcgccgc cgtacccggg tggcggcggc
      241 ggcggcggcg gtggcggcat gatgatgagt gacctcctct tctacggcgg cgacggcggc
      301 agcgcggagg cgaggggcgg gatggacgcg tcgccgttcc aggagctggc gtccatggcg
      361 gcgccgccgc ctcagcatcc gcacgaggag ttcaacttcg actgcttgag cgaggtctgc
      421 aacccgtacc gcagctgcgg cgcccagctc gtcccctcgg aggcggctag ccagacgcag
      481 acgcagctca ctcctctccg cgacgcaatg gtggcggagg aggagacgag cggcgacaag
      541 gcgctgctcc atggcggcgg cggctcgtcg tcgccgacgt tcatgtttgg aggtggcgcc
      601 ggagagagct cggagatgat ggccggcatc agaggcgtcg gcggcggcgt ccacccgcgg
      661 agcaagctcc acggcacgcc gtcgaagaac ctcatggccg agaggcggcg tcgcaagcgg
      721 ctcaacgacc ggctgtccat gctccggtcc atcgtgccca agattagcaa gatggacagg
      781 acctcgatcc ttggtgacac catcgactac gtgaaggagc tgacggagag gatcaaaacc
      841 ctcgaggagg agatcggcgt cacgccggag gagctcgacc tgttgaacac gatgaaggat
      901 tcctccagcg gcaacaacaa cgagatgctc gtgaggaatt ccaccaagtt cgatgtcgag
      961 aaccggggca gcgggaacac gaggatcgag atatgctgcc cggcgaaccc cggcgtgctg
     1021 ctgtcgacgg tgagcgcgct ggaggtgctg gggctagaga tcgagcagtg cgtcgtgagc
     1081 tgcttcagtg actttgacat gcaggcctcc tgcttacaag aggacgggaa gaggcaagtt
     1141 gtcagcaccg acgagaacaa gcagacattg ttcaggagcg caggctatgg cggaaggtgt
     1201 ctgaagggtg ggcgcgccga cccagctttc ttgtacaaag ttggcattat aagaaagcat
     1261 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1321 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     1381 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     1441 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     1501 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1561 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     1621 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1681 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     1741 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     1801 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     1861 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     1921 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     1981 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     2041 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2101 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2161 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2221 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     2281 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     2341 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     2401 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2461 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2521 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2581 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2641 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2701 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2761 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2821 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2881 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2941 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3001 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3061 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3121 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3181 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3241 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3301 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     3361 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     3421 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     3481 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     3541 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     3601 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     3661 ccc
//
