LOCUS       sequence-13196-e        4560 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 4560)
  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 4560)
  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: pUT1508-HQ858840
FEATURES             Location/Qualifiers
     source          1..4560
                     /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"
     regulatory      248..257
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2117..2216)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2233..2252)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2234..2252)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2257..2273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2257..2273)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2375..2394)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2386..3195
                     /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(2453..2472)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3342..3930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3831..3850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4084..4101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4260..4287)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4379..4465)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4514..4536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4528..4545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4529..4545
                     /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 atgaccgcca gatcgggcga ccccggcgtc tcgccggagt cgtcgtctgc ggcggcggcg
      181 gcggcggggt ccgggggcgg ggagatctgg gggacgtcgg aggacctgct cctggcgtgc
      241 gccgtgagcc gccatgggac ggcgagctgg gacgcggtgg cgaaggagat gcagtcgagg
      301 tgcccttccg ccgccgtgtt cacccccacc acctgccgcc tccggttccg cgtcctccac
      361 cgccggttct ccggcggcgt caccgccgag aacgaggacg ccgacggcgg cgaggaggag
      421 gaagaggccg acgccgccgc cgtcgccggc tgggtggagg aactccgcga gctgcgcgtc
      481 gccgagctcc gccgcgaggt cgagaaatac gatctctcga tcgggtcgtt gcaatcaaaa
      541 gtgaaacggc tcaaggagga gagggaaaag agcatctccg gcgagacgaa gccgccgccg
      601 gcggcggcgg cggtgaagga ggaggaggag gatgtacgga agggatcacc ggaggaggcc
      661 ggcgccgtgg aggacagggt ctccggccac gaatccggcc ggtcatgcaa ggagtcgaac
      721 tcgtcagatc tgaagcggcc ggagaacgac cccgcggtcg ccgacgacga cgaccgggag
      781 gcggaggagg aggaggaggc ggcggcggcg cccgcggccg gcgacatcgc cgtgaaggag
      841 gaggcgtcgg gcgagtcggt ggctgggtcg aaggaggcgg acgtcgagaa ggagagcagc
      901 gacgtgcaga gctccgccag cccgtcccgc cgccggcgac ggaagggcgg aggcggcggc
      961 gaagaggcgg aggcggcgtc gccgtcggtg tccgtgcccc tccccgccgc cgaggccgag
     1021 ccgctcgtcg ccttcctcga gtcggtccgg accagcaagg ccggcgccgt gttcgagcgg
     1081 cggctcgata gccaggatgg ggagaggtac agtggcacga tcaggcgcca tgtggacctg
     1141 gagacggtca ggtcgaggct ggtgggtgca acggcggcgg cggcggcggc ggcgtgctac
     1201 gcgtcggcgt ccgagttcta ccgcgacatg atgctgctct gcgccaacgc gctcgtcttc
     1261 ttcccgcgcg gctcgccgga gcacgccgcc gcgctccagc tgcgcgcgct cgtctccaag
     1321 caggtctcca aggaccggca gcctcacgcc ggggcgaagg cccccgcggc ggcggcggag
     1381 gaggaggaga agaagaagcc ggccaaggcc gacgccgaca tcgctgggcc gctgctcgag
     1441 aaggcgccga tcatcgtctg ccggaagcgg agctccatcg cgaaggccgc ggcggcggct
     1501 gccaaggggg agaaggcgga gaaggcagag acggacaaga aggagaagga cggcagcgag
     1561 gagaagaaga aggctgcggc ggcagcgacg acggcgacgg cggcggccac tgccaaggac
     1621 aagaaggctc ggggcatgag gacgaacaag agccggggac ccgcgaggaa tcagaagacg
     1681 gcgaagctct cggagaccgg cgaggggacc aagaagtccg acaagaaggg cggcggcgga
     1741 ggcggcagct cttcggcagg ggcggcggcg ggcggggttg ccaagaagcg aaacgccgtg
     1801 gatttcctga accggatgaa ccagaacggc tcgccgtcga cggagagggt gtcgttgctg
     1861 gagacgctga agctctccgc ggcggcgacg gagcagcaga agaagtcgtc gtcgtcgtca
     1921 tccgggaagg gcgacggccg gaaggaggcc ggcggctcag gctccaagaa gggcgccgcc
     1981 gcctcgacgc caccagggag gaggatcggt cggccaccga agcgggcggc cgcgccgccg
     2041 acgccgccgc cgtccaagag ggccaaggac gacaagccga ccaggaagcg cgggaagaag
     2101 aagggtgggc gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     2161 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     2221 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     2281 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     2341 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     2401 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2461 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     2521 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2581 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2641 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     2701 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2761 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2821 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2881 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2941 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3001 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     3061 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     3121 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     3181 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     3241 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     3301 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3361 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3421 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3481 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3541 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3601 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3661 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3721 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3781 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3841 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3901 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3961 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4021 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4081 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4141 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4201 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     4261 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     4321 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     4381 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     4441 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     4501 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
