LOCUS       sequence-13213-e        3738 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 3738)
  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 3738)
  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: pUT1620-HQ858857
FEATURES             Location/Qualifiers
     source          1..3738
                     /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(1295..1394)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1411..1430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1412..1430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1435..1451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1435..1451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1553..1572)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1564..2373
                     /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(1631..1650)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2520..3108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3009..3028
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3262..3279
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3438..3465)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3557..3643)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3692..3714
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3706..3723
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3707..3723
                     /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 catgtcttct ggaggaggag gaggaggagg aggagatcgc
      181 catggcccct accaccagca cgggcacctc ggccgcggcg aaggcgctga ttacgtgtac
      241 agcagcagcg acatggagag cttcttcttc agccaacccg ggggcgtcgg catcggcggg
      301 ggtggtggtg gcgtcgtcgg cgccggcggt gccgacgaga tcatgccgta ctccagcatc
      361 acggactacc tgcaggggtt attggacccc tccgggctag ctcggcacct cgacgtggcg
      421 tgcccgtcgt ctcaggacac ggtggtcaag caggagctgt cggtggatgt gacgagccac
      481 gacagccagg gcaccggcgg cgtcgccgga gaaggcgtcg cgcaggccac gccgaactcg
      541 tcggcgtcct tctcgtccag cgacggagag gcggagggcg gcaaatcgtc acgccggtgc
      601 aagaagggtc aggcgaaggc ggaggaggag gacgacaagg atgaggagga tggagagaac
      661 tccaagaaac cgaacaagcc caagaagaaa gctgagaaga ggcagcggca gcctcgcgtg
      721 gcgttcctca ccaagagcga ggttgatcac ctcgaggacg gctaccggtg gcgcaagtac
      781 ggccagaagg cagtcaagaa cagcccatac ccgaggagct actaccggtg cacgacgccc
      841 aagtgcggtg tgaagaagcg ggtggagcgg tcgtaccagg acccctccac ggtgatcacc
      901 acgtacgaag ggcagcacac gcaccacagc ccggcgagcc tccgcggcgg cggcggcggc
      961 gtcggcatcg tcggagggca ccaccaccac cacctcttca tgcccggcgt gcatggcctg
     1021 ccgccgtcgc acctcatgcc ggcggggttc caccctgagc tgatgggcct catgcaccac
     1081 cacccggcca tggccgccgc cgccgcaaac cctagcatgt acttcccggg cgtagctgct
     1141 tctgctccgc cgcctcctgc tgtggctggc ggcggtgcaa tgccgcccaa cgatcatcct
     1201 cctcttcagc agcatcactt cactgactac gccctgctgc aagacctttt cccttccaca
     1261 atgcccagca gcaacccaaa gggtgggcgc gccgacccag ctttcttgta caaagttggc
     1321 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1381 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1441 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1501 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1561 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1621 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1681 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1741 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1801 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1861 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1921 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1981 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2041 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2101 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2161 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2221 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2281 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2341 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2401 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2461 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2521 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2581 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2641 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2701 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2761 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2821 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2881 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2941 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3001 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3061 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3121 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3181 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3241 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3301 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3361 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3421 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3481 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3541 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3601 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3661 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3721 agtcttaagc tcgggccc
//
