LOCUS       sequence-13224-e        3762 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 3762)
  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 3762)
  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: pUT1677-HQ858868
FEATURES             Location/Qualifiers
     source          1..3762
                     /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)"
     CDS             304..321
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     protein_bind    complement(1319..1418)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1435..1454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1436..1454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1459..1475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1459..1475)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1577..1596)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1588..2397
                     /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(1655..1674)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2544..3132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3033..3052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3286..3303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3462..3489)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3581..3667)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3716..3738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3730..3747
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3731..3747
                     /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 catgaacagg atgacggcgc cgcacggggg gatgccgccg
      181 ccgccgatgc cggcggcggg ggggctggcg aggtacggct cggcgccggg gtcgctgctg
      241 gcgtcgatcg ccgactcggt gatccggggc cgcggggttg gggttgtcga tcagctgcat
      301 catcatcagc atcagcatca gcttccgccg ccgccgccgc cgcagcagca gcagatggtg
      361 gggaggtact tctccgcgga gtcgtcgggg ctcacctcct gcgagtccag ctgccggacg
      421 acgacgacga cgtccacagc ggcggcggcc gatgtcgggc ggcacccgct cgagcgcgcg
      481 tacggcggct ccggcgagat ccacgtcgac gcctcctccg ccgccgtccc gctcttccgc
      541 cacagcagct cccccgccgg cctcctctcc cgcctcatgg ctgacccgca cggcaatggc
      601 atggccgcga cgagagggat gggcggctac tccggcggcg gcggcgacgc cggcgcgatg
      661 gcgcacagga ggctgagctc gcagtggagc ttctcgcggc aggacctgcc gcagatatcg
      721 gagatgggcg gcctcatccc cgacatcggg gagagcatcg tcaccggcgg cggcggcaac
      781 agctccagca atggcgccgg ccacggcgcg cagtcctcct ccttcctctc ctccaggaac
      841 ttctccatga gctcctggga cgacaccaac tccatcatgt tctcgccccc gagcagcagc
      901 aagaaggccc gcgtcgccgc cgccgccgcc ggcgaccacg gcgacgacat ggtctccagc
      961 ttcagcaaca tcgactccca gtttggcttg tcgaagcagt cgtcgctgga gatggccggc
     1021 atggatgact tcttgcagct ccagccagac tccgtcgcct gcagggcccg ggcaaagcgc
     1081 ggctgcgcga cgcacccgcg gagcatcgct gagagggaga ggagaacgag gattagcaag
     1141 aggctgaaga agctgcagga tctcgtgccg aacatggaca agcaaacgaa tacatcagac
     1201 atgttggata ttgctgtaac ctacatcaag gagcttcagg gccaagttga gaagctaaaa
     1261 catgaccaag caaactgcac ttgctcaggc aagcacgatt gcaagggtgg gcgcgccgac
     1321 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     1381 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     1441 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1501 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     1561 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     1621 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1681 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     1741 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1801 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1861 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1921 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1981 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2041 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2101 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2161 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2221 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2281 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2341 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     2401 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     2461 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2521 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2581 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2641 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     2701 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2761 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2821 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2881 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     2941 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3001 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3061 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3301 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3361 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3421 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3481 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3541 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3601 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3661 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     3721 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
