LOCUS       sequence-160249-        4278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway donor vector for AFB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4278)
  AUTHORS   Havens KA, Guseman JM, Jang SS, Pierre-Jerome E, Bolten N, Klavins 
            E, Nemhauser JL
  TITLE     A synthetic approach reveals extensive tunability of auxin 
            signaling.
  JOURNAL   Plant Physiol. 2012 Sep;160(1):135-42. doi: 10.1104/pp.112.202184. 
            Epub 2012 Jul 27.
  PUBMED    22843664
REFERENCE   2  (bases 1 to 4278)
  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.112.202184"; journalName: "Plant Physiol"; date: 
            "2012-09"; volume: "160"; issue: "1"; pages: "135-42"
COMMENT     Sequence Label: pDONR-AFB2
FEATURES             Location/Qualifiers
     source          1..4278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2403..2502)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2519..2538)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2520..2538)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2543..2559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2543..2559)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2661..2680)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2672..3481
                     /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(2739..2758)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3628..4216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4117..4136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc aaaatgaatt atttcccaga tgaagtaata gagcatgtat tcgactttgt
      721 aacatctcac aaagacagga atgctatatc tcttgtatgc aaatcatggt acaagattga
      781 aagatacagt aggcaaaagg ttttcattgg aaactgttat gccattaatc cagagaggtt
      841 gcttcggaga ttcccatgtc taaagtcttt gactttgaaa ggaaaacctc attttgcgga
      901 tttcaatttg gttcctcatg aatggggagg ttttgtgcta ccttggattg aggctttggc
      961 tagaagccgt gtaggacttg aagagcttag gttgaagagg atggttgtta ctgatgagag
     1021 tcttgagctg ctttctcgtt cttttgtcaa ttttaagtct ttggtccttg ttagctgtga
     1081 aggttttacc actgatggtc ttgcctctat tgccgctaat tgcaggcatc ttcgggatct
     1141 tgatttgcaa gagaatgaaa tcgatgatca tagaggtcaa tggttaagtt gtttcccaga
     1201 cacttgcacg actcttgtca cgctaaactt tgcgtgcctc gaaggagaaa ctaatctggt
     1261 tgctctagag aggcttgttg ctaggtctcc aaacctaaag agtctgaagc taaatcgtgc
     1321 agtaccgtta gatgcactcg caaggttaat ggcgtgtgcg ccgcagatag ttgacttagg
     1381 agtagggtct tatgagaatg acccagattc cgagtcttac ttgaaactca tggctgtcat
     1441 aaagaaatgc acctcgttga ggagtttgtc gggttttcta gaggctgctc ctcactgtct
     1501 ctcagctttc cacccaatat gtcataacct cacctccttg aatcttagtt acgcagctga
     1561 gattcatggt agccacctta ttaagcttat tcagcattgc aagaaacttc agcggttatg
     1621 gattttggat agtataggtg acaaagggct tgaagttgta gcttctacat gtaaagagtt
     1681 acaagagctt agggtttttc catctgattt actcggtgga ggcaacacag ctgtgaccga
     1741 agaaggtcta gttgccatct cggcaggctg ccctaagctc cactctatac tctacttctg
     1801 ccaacaaatg acaaacgcag ctctcgtaac cgttgccaag aactgtccaa atttcatccg
     1861 tttccgactc tgcatcctcg agccaaacaa acccgatcac gtcacatctc aacctctaga
     1921 cgaaggcttt ggagcaatcg tcaaagcctg caagagcctg agaaggcttt ctctctcagg
     1981 tctccttaca gaccaagtct tcctctacat cggaatgtac gcgaatcagc tcgagatgct
     2041 ctccatagcc tttgcaggag atacagacaa aggcatgcta tatgtgttga atggttgcaa
     2101 aaagatgaag aaactagaga taagggatag tccgtttggg gacacggcgc ttcttgctga
     2161 tgtgagcaag tatgaaacaa tgcgatccct ttggatgtct tcatgtgaag tcacactcag
     2221 tggatgcaaa aggctcgcag agaaagcgcc atggctcaat gtagagatca taaacgagaa
     2281 tgataataac cggatggaag aaaacggaca cgaggggagg cagaaagtgg ataagttgta
     2341 tctgtaccgg actgtggttg ggacaagaat ggatgcgccg ccatttgtgt ggattctcta
     2401 acacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2461 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2521 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2581 aaatctctga tgttacattg cacaagataa aataatatca tcatgaacaa taaaactgtc
     2641 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2701 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2761 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2821 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2881 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2941 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     3001 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     3061 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3121 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3181 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3241 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3301 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3361 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3421 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3481 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3541 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4261 gccttttgct cacatgtt
//
