LOCUS       sequence-13183-e        3333 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 3333)
  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 3333)
  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: pUT1325-HQ858827
FEATURES             Location/Qualifiers
     source          1..3333
                     /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(890..989)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1006..1025)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1007..1025)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1030..1046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1030..1046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1148..1167)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1159..1968
                     /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(1226..1245)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2115..2703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2604..2623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2857..2874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3033..3060)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3152..3238)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3287..3309
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3301..3318
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3302..3318
                     /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 atggggaggt caccgtgctg cgagaaggca cacaccaaca agggagcatg gaccaaggag
      181 gaagatgacc ggctcattgc ctacatcaag gcgcacggcg aaggttgctg gcgatcgctg
      241 cccaaggccg ccggcctcct ccgctgtggc aagagctgcc gcctccggtg gatcaactac
      301 ctccggcctg acctcaagcg cggcaacttc accgaggagg aggatgagct gatcatcaag
      361 cttcacagcc ttttaggcaa caaatggtct ctgatagccg ggaggttgcc aggaagaacg
      421 gacaacgaga tcaagaacta ctggaacacg cacatcagga ggaagctgct gagccgtggc
      481 atcgacccgg tgacacaccg gccgatcaac gacagcgcgt ccaacatcac catatcattc
      541 gaggcggccg cggcggcggc gagggacgac aaggccgccg tgttccggcg agaggaccat
      601 cctcatcagc cgaaggcggt gacagtggca caggagcagc aggcagccgc cgattggggc
      661 catgggaagc cactcaagtg ccctgacctc aatctggacc tctgcatcag cctcccttcc
      721 caagaagagc ccatgatgat gaagccggtg aagagggaga ccggcgtctg cttcagctgc
      781 agcctggggc tccccaagag cacagactgc aagtgcagca gcttcctggg actcaggaca
      841 gccatgctcg acttcagaag cttggaaatg aaaaagggtg ggcgcgccga cccagctttc
      901 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
      961 actatcagtc aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg
     1021 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     1081 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1141 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1201 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1261 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1321 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1381 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1441 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     1501 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1561 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1621 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1681 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     1741 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     1801 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     1861 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     1921 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     1981 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     2041 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2881 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2941 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     3001 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3061 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     3121 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     3181 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     3241 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     3301 tgtaaaacga cggccagtct taagctcggg ccc
//
