LOCUS       sequence-13191-e        3537 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 3537)
  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 3537)
  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: pUT1345-HQ858835
FEATURES             Location/Qualifiers
     source          1..3537
                     /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"
     regulatory      985..994
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1094..1193)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1210..1229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1211..1229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1234..1250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1234..1250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1352..1371)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1363..2172
                     /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(1430..1449)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2319..2907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2808..2827
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3061..3078
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3237..3264)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3356..3442)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3491..3513
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3505..3522
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3506..3522
                     /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 atggaaggcg ttgttgacgg gagcggcgcg cagctggtgg tggcggagct ggtgcgcgtg
      181 caggggctgc tccggcagct cgaggcccac ctgagcgcgc cgtgctccgt cgagctctgc
      241 cgcggcctcg tcgcgcagat cgtcgcgctc acggacaggt ccatcggcat cgccacccgg
      301 tccttctcct ccgcctccgg cggcggcgcg catttcgcgg acacggcgcc gccgatgccg
      361 gcgctgacgt cgtgcacgcc cagcccgctc agcgacggct ccgaccacca gccgttccgg
      421 accaccaacg ccaagaagag gaagacgacg gcgaggtgga cgagccaggt gagggtgagc
      481 gcggcgggcg gcgcggaggg gccagccgac gacgggcaca gctggaggaa gtacgggcag
      541 aaggacatcc tgggcgccaa gcacccgcgc ggctactacc gctgcaccca ccgcaacacg
      601 cagggatgca ccgccaccaa gcaggtccag cgcaccgacg acgacgcctc cctcttcgac
      661 gtcgtctacc acggcgagca cacctgccgc ccgggcgccg ccagcgccgc cgccgccaag
      721 aggccgcacg cgcagacgct gctgcagagc ctcagcgcca gcctcaccgt gaacacggac
      781 accaacacgc cgctgacgcc ggagaaccgc gcgccggcgc cgccgctgca gcagcagcag
      841 cagcagcaga gcgtgtcggc gtcgcccgtg gcgtcggact cgtacggcct gggcggcgcc
      901 gggtacgggg actggcgctg ctgcgacggc gacctgcagg aggtcgtgtc ggcgctcgcc
      961 accgtgacgt cggcccccga ccacgccgcc atggacgccg ccgacttcat gagctactgc
     1021 ttcgacttcg acccggcggt gtacggcggc atcgtcggca cgccgagctt cttcctaaag
     1081 ggtgggcgcg ccgacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1141 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1201 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1261 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     1321 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1381 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1441 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     1501 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1561 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1621 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     1681 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1741 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1801 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1861 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     1921 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1981 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2041 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2101 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2161 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2221 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     2281 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2341 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2401 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2461 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2521 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2581 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2641 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2701 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2761 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2821 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2881 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2941 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3001 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3061 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3121 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3181 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3241 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     3301 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     3361 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     3421 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     3481 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
