LOCUS       40924_29936        2115 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid with a Zeocin(TM) resistance marker and a TEV 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 2115)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 2115)
  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..2115
                     /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=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQS"
     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             complement(938..1309)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1328..1375)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     rep_origin      1465..2053
                     /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 gagaacctgt acttccaatc 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 atgatcagtc ctgctcctcg gccacgaagt
      961 gcacgcagtt gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga
     1021 tctcggtcat ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact
     1081 cggcgtacag ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca
     1141 cctggtcctg gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt
     1201 cgtcctccac gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg
     1261 cgacgtcgcg cgcggtgagc accggaacgg cactggtcaa cttggccatg gtttagttcc
     1321 tcaccttgtc gtattatact atgccgatat actatgccga tgattaattg tcaacacgtg
     1381 ctgatcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     1441 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1501 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1561 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1621 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1681 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1741 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1801 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1861 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1921 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1981 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2041 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2101 ttttgctcac atgtt
//
