pENTR3C-Dual

LOCUS       Exported                3756 bp ds-DNA     circular SYN 13-AUG-2016
KEYWORDS    pENTR3C-Dual
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3756)
  AUTHORS   Biofeng Lab.
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3756
                     /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"
     promoter        526..556
                     /note="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             610..1290
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             1610..1915
                     /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"
     protein_bind    complement(1985..2084)
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2207..3016
                     /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      3106..3694
                     /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 gcaggctctt taaaggaacc aattcagtcg
      481 actggatccg gtaccgaatt cgcggccgca ttaggcaccc caggctttac actttatgct
      541 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
      601 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
      661 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
      721 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
      781 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
      841 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
      901 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
      961 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1021 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     1081 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     1141 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1201 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcaggcg
     1261 gggcgtaatc tagaggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     1321 ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag
     1381 gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa
     1441 ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt
     1501 cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg
     1561 tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa
     1621 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     1681 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     1741 taaagtcccc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     1801 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     1861 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     1921 aggctccctt atacacagcc agtctgcagg tcgacgaatt cgcggccgca ctcgagatat
     1981 ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2041 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcagct
     2101 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2161 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2221 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2281 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2341 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2401 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2461 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2521 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2581 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2641 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2701 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2761 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2821 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2881 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2941 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3001 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cattattcag attgggcccc
     3061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3241 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3721 gttcctggcc ttttgctggc cttttgctca catgtt
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