LOCUS       sequence-13201-e        3981 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 3981)
  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 3981)
  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: pUT1549-HQ858845
FEATURES             Location/Qualifiers
     source          1..3981
                     /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             456..464
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(1538..1637)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1654..1673)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1655..1673)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1678..1694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1678..1694)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1796..1815)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1807..2616
                     /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(1874..1893)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2763..3351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3252..3271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3505..3522
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3681..3708)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3800..3886)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3935..3957
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3949..3966
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3950..3966
                     /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 ccccttcacc
      121 atgagcgcgg cggcggagga ggcggctgcg gcggcggcgg cggaggaggt ggagggggtg
      181 gaggcgggga agaagggcgt ggcggatgcg aaggggaagg ggaaggagaa ggtggtggtg
      241 gcggaggagg attctgcggt ggcggcgggc tccgggggaa gcggaggcgg gaggaggacg
      301 ttggtggcgt acccggcgcg ggttgcgggg tacaaggacg tggtggcgga cgcggcggtc
      361 ttcaggaggg cgctcgaggg gttgcacgcc cagatgggga caaagctcaa ggtgccaatt
      421 attggtggaa aagaccttga tcttcatcag ctgtttaagg aggtcacttc acgaggtggc
      481 attgataagg tgaagtccga taatagatgg agggaagtaa cagcatcatt tattttccct
      541 gctactgcga caaatgcttc ttttatgttg aagaaatact acatgtcact attataccat
      601 tttgagaggc tatacttgtt tgaggcacaa ggctggtatc aagaaactga ttccagaagc
      661 atttcctgta tagaaatgaa agctgaaggt caagcttccc ggaaaaggaa aagaggaagc
      721 aattcttgtt cttcagacct agctgcttct ttggataatg atgtacaagt gataattgat
      781 ggtaagtttg aacatggcta catagttacc gttatcatgg gctcaaaatc aacgaaagcg
      841 gtactctata attgcaccga agaacctgca gtgccaactg cggtgcctca tgttgcaatt
      901 gacagcgctg agggcatacg tcctcgccgc cgccgccgta ggaagaagtt aagtacaaca
      961 gaccccaacc atcctaaacc aaacaggagc ggctacaatt tcttcttcca agatcagcac
     1021 aggaagctca agccagaata tcctggacaa gacaggctta tcagcaagat gattggtgaa
     1081 cgatggaaca atttaggccc tgaagataaa gcagtatatc aagaaaaagg tgtagaggac
     1141 aaggcaaggt accagagaca gttagctctt tacagagaac agagaacagg ccagccaatc
     1201 agcaacgcgg tgcctatcca gcagagactt cctcagaaag aagtgacgat tgatgaggtg
     1261 gactctaaag tcagtgaagg tgatatcctg ctgtccaacc aaggatacag cagcagtacc
     1321 agtagcagtg atgaaaccgc tgattcggga gaaaaaaatg tggaagatga tgaagagttt
     1381 aacacagaga catctcctga gccgagcatg gaaacaacag attctcatgg acagcctgat
     1441 ccttctgctg atggggagcg ctttgaactg cgccgcaggg agaaccctaa gattgatgag
     1501 aaacgtgaca tgccacctaa taagggtggg cgcgccgacc cagctttctt gtacaaagtt
     1561 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     1621 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     1681 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     1741 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     1801 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     1861 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1921 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1981 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2041 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2101 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2161 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2221 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2281 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     2341 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     2401 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     2461 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     2521 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     2581 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     2641 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     2701 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2761 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2821 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2881 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     2941 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3001 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3061 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3121 acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa
     3181 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3241 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3301 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3361 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3421 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3481 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3541 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3601 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
     3661 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
     3721 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     3781 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     3841 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     3901 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
     3961 gccagtctta agctcgggcc c
//
