LOCUS       40924_29946        2586 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid with a kanamycin resistance marker and a TVMV cleavage site,
            for creating a Gateway(R) entry vector by ligation-independent 
            cloning (LIC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2586)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 2586)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For LIC, linearize with SspI and treat with T4 DNA polymerase plus 
            dGTP.
FEATURES             Location/Qualifiers
     source          1..2586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(387..473)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             671..691
                     /codon_start=1
                     /label=TVMV site
                     /note="tobacco vein mottling virus (TVMV) NIa protease 
                     recognition and cleavage site"
                     /translation="ETVRFQS"
     protein_bind    complement(711..810)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(828..846)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(851..867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             980..1786
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1936..2524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gagaccgtgc gcttccaatc caatattgga agtggataac acccagcttt
      721 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
      781 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
      841 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
      901 ttacattgca caagataaaa taatatcatc atgaacaata aaactgtctg cttacataaa
      961 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1021 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1081 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1141 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1201 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1261 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1321 aggtgaaaac attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1381 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1441 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1501 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     1561 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     1621 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     1681 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     1741 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     1801 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     1861 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     1921 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1981 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2041 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2101 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2161 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2221 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2281 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2341 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2401 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2461 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2521 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2581 catgtt
//
