LOCUS       sequence-13200-e        3330 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 3330)
  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 3330)
  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: pUT1546-HQ858844
FEATURES             Location/Qualifiers
     source          1..3330
                     /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"
     protein_bind    complement(887..986)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1003..1022)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1004..1022)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1027..1043)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1027..1043)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1145..1164)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1156..1965
                     /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(1223..1242)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2112..2700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2601..2620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2854..2871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3030..3057)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3149..3235)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3284..3306
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3298..3315
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3299..3315
                     /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 atgaaggtgc agtgcgacgt gtgcgcggcc gaggccgcct cggtcttctg ctgcgccgac
      181 gaggccgcgc tgtgcgacgc gtgcgaccgc cgcgtccaca gcgcgaacaa gctcgccggg
      241 aagcaccgcc gattctccct cctccaaccg ttggcgtcgt cgtcgtccgc ccagaagcca
      301 ccgctctgcg acatctgtca ggagaagagg gggttcttgt tctgcaagga ggacagggcg
      361 atcctgtgcc gggagtgcga cgtcacggtg cacaccacga gcgagctgac gaggcggcac
      421 ggccggttcc tcctcaccgg cgtgcgcctc tcgtcggcgc cgatggactc ccccgcgccg
      481 tcggaggaag aggaggagga agcaggggag gactacagct gcagccccag cagcgtcgcc
      541 ggcaccgccg cggggagcgc gagcgacggg agcagcatct ccgagtacct caccaagacg
      601 ctgcccggtt ggcacgtcga ggacttcctc gtcgacgagg ccaccgccgc ctcctcctcc
      661 tcagacgggc tatttcaggg tgggctgctg gctcagatcg gtggggtgcc ggacggttac
      721 gcggcgtggg ccggccggga gcagctgcac agtggcgtcg ctgtcgccgc cgacgagcgg
      781 gccagccgcg agcggtgggt gccgcagatg aacgcggagt ggggcgccgg cagcaagcga
      841 cccagggcgt cgcctccctg cttgtactgg aagggtgggc gcgccgaccc agctttcttg
      901 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
      961 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     1021 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     1081 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1141 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     1201 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     1261 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     1321 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     1381 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     1441 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     1501 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     1561 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     1621 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     1681 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     1741 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     1801 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     1861 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     1921 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     1981 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     2041 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     2101 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2161 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2221 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2281 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2341 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2401 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2461 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     2521 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2581 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2641 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2701 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2761 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2821 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2881 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2941 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     3001 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     3061 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     3121 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     3181 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     3241 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     3301 aaaacgacgg ccagtcttaa gctcgggccc
//
