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LOCUS       Exported                4773 bp ds-DNA     circular SYN 10-AUG-2016 .
KEYWORDS    pDONR-P2RP3
REFERENCE   1  (bases 1 to 4773)
  AUTHORS   Biofeng Lab.
FEATURES             Location/Qualifiers
     source          1..4773
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(591..822)
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP2"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     promoter        980..1082
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1083..1742
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2084..2389
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2746..2977)
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP3"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3015..3033)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3038..3054)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3167..3976
                     /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      4123..4711
                     /direction=RIGHT
                     /note="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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg attcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagttggca ttataaaaaa gcattgctta tcaatttgtt
      781 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttggagctct agagcgtcga
      841 ctaagttggc agcatcaccc gacgcacttt gcgccgaata aatacctgtg acggaagatc
      901 acttcgcaga ataaataaat cctggtgtcc ctgttgatac cgggaagccc tgggccaact
      961 tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac cataatgaaa
     1021 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     1081 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     1141 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1201 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     1261 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     1321 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     1381 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     1441 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     1501 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     1561 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     1621 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc
     1681 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     1741 aatcgcgtgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     1801 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtgtgc
     1861 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     1921 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     1981 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     2041 gaaatgaacg gctcttttgc tgacgagaac agggactggt gaaatgcagt ttaaggttta
     2101 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     2161 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     2221 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     2281 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     2341 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     2401 ccttatacac agccagtctg caggtcgata cagtagaaat tacagaaact ttatcacgtt
     2461 tagtaagtat agaggctgaa aatccagatg aagccgaacg acttgtaaga gaaaagtata
     2521 agagttgtga aattgttctt gatgcagatg attttcagga ctatgacact agcgtatatg
     2581 aataggtaga tgtttttatt ttgtcacaca aaaaagaggc tcgcacctct ttttcttatt
     2641 tctttttatg atttaatacg gcattgagga caatagcgag taggctggat acgacgattc
     2701 cgtttgagaa gaacatttgg aaggctgtcg gtcgagctcg aattctacag gtcactaata
     2761 ccatctaagt agttgattca tagtgactgc atatgttgtg ttttacagta ttatgtagtc
     2821 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt
     2881 tcaactttat tatacaaagt tggcattata aaaaagcatt gcttatcaat ttgttgcaac
     2941 gaacaggtca ctatcagtca aaataaaatc attatttgga gctccatggt agcgttaacg
     3001 cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     3061 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     3121 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     3181 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     3241 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     3301 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     3361 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     3421 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     3481 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3541 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3601 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3661 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3721 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3781 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3841 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3901 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3961 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     4021 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     4081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4261 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4501 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4741 cctggccttt tgctggcctt ttgctcacat gtt
//

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