LOCUS       sequence-13206-e        3630 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 3630)
  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 3630)
  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: pUT1613-HQ858850
FEATURES             Location/Qualifiers
     source          1..3630
                     /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)"
     protein_bind    complement(1187..1286)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1303..1322)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1304..1322)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1327..1343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1327..1343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1445..1464)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1456..2265
                     /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(1523..1542)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2412..3000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2901..2920
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3154..3171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3330..3357)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3449..3535)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3584..3606
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3598..3615
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3599..3615
                     /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 catggatccg cgcagcagcc aggcgcaaga ggacggcttc
      181 ttccacccac gcgacggcgc gtgccccgcc gactccagcg gaaagacaga atgtaaaacc
      241 cagggttcaa ttgccacgcg caaggttcaa aaggctgatc gtgagaaaat gcgaagagat
      301 agattgaatg agcagttcca ggaacttgga agcacacttg atccagatcg acccaggaat
      361 gataaggcaa ctatccttag tgatgcaatt caaatgctga aagatttgac gtcccaggtg
      421 aacaagctta aagctgaata tacttcactc tcagaagaag cacgtgagtt gactcaggaa
      481 aagaatgagc tcagagatga gaaagtttca ctgaagtttg aggttgataa tctgaatacc
      541 cagtatcagc aacgaatgcg tgtgctgtac ccttggactg gaatggagcc ttctgttgtc
      601 attgggccac ccctgccata tcctttttca gtcccagtcc cagtcccagt ccctatacct
      661 tcaggtgctg ttcctatgca cccacagctc caggcatacc cttacttccg aaatcaaact
      721 tctggaaccg tctccaatcc atgcacacct tacatggcat acactcaacc catccatcct
      781 cctactgatc aactatctaa ccaattcagt gctccagttc agcattcaag tagtaatcga
      841 tctcactcta tggcccaaga ctctagaagc aagtcctctg cactgcagca agtaagttgt
      901 agagggaaac atgatgactt tgatgacgtt gcaactgatc tggaactaaa aactcctggt
      961 tcctcagccc ctctacaatc agagattgcc aacaaggatt cctcttctga cttgaaaaaa
     1021 aagcagcagt tcatacagga aactaaaggt agttctctta cagaaggcag tagtagttca
     1081 agcagatgct cgtccagtgg tcctccagat gtctctaaca gtattgaagg tggatcagtt
     1141 gctgatgatc aacgatcgac agtacagaca aagggtgggc gcgccgaccc agctttcttg
     1201 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1261 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     1321 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     1381 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1441 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     1501 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     1561 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     1621 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     1681 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     1741 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     1801 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     1861 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     1921 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     1981 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2041 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     2101 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     2161 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     2221 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     2281 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     2341 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     2401 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2461 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2521 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2581 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2641 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2701 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2761 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     2821 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2881 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2941 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3001 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3061 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     3301 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     3361 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     3421 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     3481 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     3541 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     3601 aaaacgacgg ccagtcttaa gctcgggccc
//
