LOCUS       sequence-13217-e        3690 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 3690)
  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 3690)
  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: pUT1670-HQ858861
FEATURES             Location/Qualifiers
     source          1..3690
                     /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)"
     regulatory      197..206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1247..1346)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1363..1382)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1364..1382)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1387..1403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1387..1403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1505..1524)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1516..2325
                     /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(1583..1602)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2472..3060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2961..2980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3214..3231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3390..3417)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3509..3595)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3644..3666
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3658..3675
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3659..3675
                     /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 catggcgttg gaggcggtgg tgttctcgga aggatactac
      181 ttcggatgcg gcggtgccat ggcggcggaa gcagctgcag gaggagcttg gagctggagc
      241 cacgggtatg gcggtggcgt tgatcagggg aagggggcga tggagttagt ggtggacgac
      301 ggggtggtga acgccttctg ggacggcggc ggaggcggcg caagctcttc cccggtgatg
      361 gccgccgtct cggggtttat tgaggagccg gacggcggta attcctccgt cgcagacgca
      421 gcgccgccgc cggagcacgg aggtgcgtct gtcgtcggac gggatggctc cgcggcggcg
      481 ccggcggcgg ccgcagggag gaggaagcgg cggagggcga ggaccgtcaa gaacagggag
      541 gaggtggaga gccagcgcat gacccacatc gccgtcgagc gcaaccgccg gaagcagatg
      601 aacgagtacc tcgccgtgct ccgctccctc atgccggcct cctacgtgca aaggggcgac
      661 caggcgtcca taattggcgg agcaatcaac tacgtcaagg agatggagca gcttctccaa
      721 tccctggagg cgcacagaca cgcccgccgc gcacgcaccg acgccgccgc cgcgttgcca
      781 ttcgcgggct tcttcacgtt cccgcagtac tccatgagcg ccgtgcccac gacaaccacc
      841 accactgtag cagcagcagc aacggaaaac ggcaatgccg tcgccggcgc cggcgcgggc
      901 gacgacgcgg acgcggacgt gtcggggtcg aagccgtcct cggtggcgga catcgaggtg
      961 acgatggtgg agagccacgc caacctgaag gtgctgtcgc ggcggcggcc gaggcagctg
     1021 ctgaggatgg tggcagggct gcaacaccac cgcctcgccg tgctccacct caacgtcgcc
     1081 agcgccggcc acatggctct ctactccctt agccttaagg tggaagaaga ctgccagctc
     1141 acgtcggtgg acgacatcgc ggcggcggta cacgggatcg tcgagacgat cgagcaagag
     1201 cagcagcagc agcagaagca gagctgtagc aagggtgggc gcgccgaccc agctttcttg
     1261 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1321 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     1381 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     1441 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1501 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     1561 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     1621 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     1681 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     1741 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     1801 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     1861 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     1921 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     1981 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     2041 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     2101 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     2161 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     2221 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     2281 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     2341 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     2401 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     2461 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2521 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2581 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2641 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2701 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2761 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2821 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     2881 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2941 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3001 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3061 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3121 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3181 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3241 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3301 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     3361 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     3421 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     3481 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     3541 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     3601 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     3661 aaaacgacgg ccagtcttaa gctcgggccc
//
