LOCUS       Exported                3781 bp ds-DNA     circular SYN 13-AUG-2016
KEYWORDS    pENTR11-Dual
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3781)
  AUTHORS   Biofeng Lab.
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3781
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL1"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(567..872)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /note="ccdB"
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1192..1872)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     promoter        complement(1926..1956)
                     /note="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(2010..2109)
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2232..3041
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3131..3719
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttcg aaggagatag aaccaattct
      481 ctaaggaaat acttaaccat ggtcgactgg atccggtacc gaattcgtcg acctgcagac
      541 tggctgtgta taagggagcc tgacatttat attccccaga acatcaggtt aatggcgttt
      601 ttgatgtcat tttcgcggtg gctgagatca gccacttctt ccccgataac ggagaccggc
      661 acactggcca tatcggtggt catcatgcgc cagctttcat ccccgatatg caccaccggg
      721 taaagttcac gggggacttt atctgacagc agacgtgcac tggccagggg gatcaccatc
      781 cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca caaacagacg ataacggctc
      841 tctcttttat aggtgtaaac cttaaactgc atttcaccag cccctgttct cgtcagcaaa
      901 agagccgttc atttcaataa accgggcgac ctcagccatc ccttcctgat tttccgcttt
      961 ccagcgttcg gcacgcagac gacgggcttc attctgcatg gttgtgctta ccagaccgga
     1021 gatattgaca tcatatatgc cttgagcaac tgatagctgt cgctgtcaac tgtcactgta
     1081 atacgctgct tcatagcata cctctttttg acatacttcg ggtatacata tcagtatata
     1141 ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg ttatctggct tttagtaagc
     1201 cggatcctct agattacgcc ccgcctgcca ctcatcgcag tactgttgta attcattaag
     1261 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     1321 cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt
     1381 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac
     1441 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     1501 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     1561 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     1621 tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg
     1681 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     1741 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
     1801 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     1861 ttttttctcc attttagctt ccttagctcc tgaaaatctc gacggatcct aactcaaaat
     1921 ccacacatta tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgcggccgc
     1981 gaattcgcgg ccgcactcga gatatctaga cccagctttc ttgtacaaag ttggcattat
     2041 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2101 cattatttgc catccagctg cagctctggc ccgtgtctca aaatctctga tgttacattg
     2161 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2221 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     2281 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     2341 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2401 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2461 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2521 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2581 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2641 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     2701 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2761 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     2821 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2881 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2941 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     3001 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     3061 tgtaacatta ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca
     3121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3241 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3781 t
//