LOCUS       sequence-13204-e        3600 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 3600)
  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 3600)
  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: pUT1611-HQ858848
FEATURES             Location/Qualifiers
     source          1..3600
                     /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(1157..1256)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1273..1292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1274..1292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1297..1313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1297..1313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1415..1434)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1426..2235
                     /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(1493..1512)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2382..2970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2871..2890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3124..3141
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3300..3327)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3419..3505)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3554..3576
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3569..3585
                     /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 catggcgtct cctgatggtg gcgttggaga cggcggcgcg
      181 gagccacatg aagttatgga cgacctgctc gagatgcgcg agcaagcggc gatgctccac
      241 agcatgctac atgggacatc gccgtcgagc tgtgcggcgg cggcgagcac caggcagctg
      301 aatcagctga tagacggggt gatgagcagg ctgcagtcca gctcgctgtc ggtgatgtcg
      361 cctggaggag gaggaggaag aagaggatcg ggcgggagga agaagaaagg cgccaaggcc
      421 gtggccgggc cgcacaggcg gagcagctcc ggtagaagaa gatcgaaaag tccttttgtc
      481 aggatggtca cgaccaagga gctggaggac ggcaggcaat ggcgaaagta cgggcagaaa
      541 catatacagg actccccaaa taatccaagg agctactaca ggtgcacgca taggccagac
      601 caaggttgta tggccacaaa gcaagtacag acctccgagt ctaactcgtc ggagttcgtc
      661 atcagctact acggtgagca cacctgcagc gacccttcca ccatcccgtt cgtcgtcgag
      721 gccgaagctc ccgctgcaga ctacgcgaac ctcatcagct tcggatccag cggcggcgcg
      781 agcacaagca gagtcgaccc tcttcgtcag agccgccatc gtttgatggc agaagccgtt
      841 gatccgacgc cgtcctgcag cttcgccaac tgccactctc cggtactgtc ctcggagtgc
      901 gccagcgagg cggcggcgct cagcagcagc ctgccgctct ccgccgtggt gggatcggcg
      961 gtgacaacgc cgtcgacgtc gatcgtgggt tcagcgcctg cggattacga ctggccgtca
     1021 gggctcgccg gaggtgacat ggccggcagc ttcccgtcct ctcccagcag cctcgggttc
     1081 atgactggct cctttgggaa tcttcctggg gatgacgacg acatgtttgg cttcgatcct
     1141 aagggtgggc gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     1201 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1261 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     1321 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1381 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1441 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1501 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1561 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1621 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1681 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     1741 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     1801 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1861 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     1921 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     1981 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2041 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2101 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2161 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2221 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     2281 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     2341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2521 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2761 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2821 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2881 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2941 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3001 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3061 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3121 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3181 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3241 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     3301 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     3361 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     3421 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     3481 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     3541 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
