LOCUS       sequence-13233-e        3438 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 3438)
  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 3438)
  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: pUT1755-HQ858877
FEATURES             Location/Qualifiers
     source          1..3438
                     /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      140..149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(995..1094)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1111..1130)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1112..1130)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1135..1151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1135..1151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1253..1272)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1264..2073
                     /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(1331..1350)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2220..2808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2709..2728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2962..2979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3138..3165)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3257..3343)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3392..3414
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3406..3423
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3407..3423
                     /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 catggcggcg gaggccggca gcgcggccgc cggcgccgcc
      181 tacgaggagg agcgccggaa gcgggtcctc gagaacctca agcacctcga ggatctgggc
      241 ataaagaaga tgtccaagag cttgctggag gcggcgagat tgcagaagag cactcgtgcc
      301 agccccaagc cgcggaagaa gtttgaggtt ggggccactg aagtgaggcg ttcatcgagg
      361 gcgcgcaact cggtttcgta caaagagaat tttgatgaac tcaacagttt tctatgccgt
      421 agaaggggca gtaggattag gagcacagag caggggaggg attacaccgg aagagttgcc
      481 tcttatgaac agcaacaacg tgctttcaaa aaagctgaga gacttcaaaa tagtctagac
      541 cctgagaacc cgtcgtttgt taagaccatg gttcgatcac atgtgtccag ctgcttttgg
      601 cttggtcttc gaactagatt ctgcaaattg catctgcctc ccaaagagta taagatggtc
      661 ttggaggatg aagagggtgg tgaatttgat tctgtctaca ttgggaaccg tacaggccta
      721 agtggtggat ggagaggttt tgcgatgcac cacaacttgg aagatggtga ttcattggtc
      781 tttgaattgg ctgaaccaga cagatttaag atctacatca taaaagctgt tgatgaggat
      841 gcaaatgagt ctgagcctgc tgatgaggaa gctattgggg acaaggacac cagtacagag
      901 gatgcagctg agcaggatga ctcacctaat gctgaacctt taaaaggaac taagaggaga
      961 aaactccgtg gacgccggaa gggtgggcgc gccgacccag ctttcttgta caaagttggc
     1021 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1081 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1141 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1201 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1261 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1321 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1381 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1441 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1501 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1561 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1621 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1681 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1741 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1801 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1861 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1921 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     1981 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2041 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2101 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2161 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2341 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2581 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3001 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3061 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3121 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3181 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3241 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3301 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3361 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3421 agtcttaagc tcgggccc
//
