pDONR221-P4RP3R

LOCUS       Exported                4717 bp ds-DNA     circular SYN 10-AUG-2016
KEYWORDS    pDONR221-P4rP3r
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4717)
  AUTHORS   Biofeng Lab.
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4717
                     /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(570..801)
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP4"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1149..1454)
                     /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"
     CDS             complement(1804..2457)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     promoter        complement(2458..2560)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    2705..2936
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP3"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2959..2977)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2982..2998)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3111..3920
                     /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      4067..4655
                     /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 tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
      781 gtcaaaataa aatcattatt tgtcgaccga cagccttcca aatgttcttc tcaaacggaa
      841 tcgtcgtatc cagcctactc gctattgtcc tcaatgccgt attaaatcat aaaaagaaat
      901 aagaaaaaga ggtgcgagcc tcttttttgt gtgacaaaat aaaaacatct acctattcat
      961 atacgctagt gtcatagtcc tgaaaatcat ctgcatcaag aacaatttca caactcttat
     1021 acttttctct tacaagtcgt tcggcttcat ctggattttc agcctctata cttactaaac
     1081 gtgataaagt ttctgtaatt tctactgtat cgacctgcag actggctgtg tataagggag
     1141 cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg
     1201 tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg
     1261 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact
     1321 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca
     1381 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa
     1441 accttaaact gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat
     1501 aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag
     1561 acgacgggct tcattctgca tggttgtgct taccagaccg gagatattga catcatatat
     1621 gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca
     1681 tacctctttt tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat
     1741 cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcca cgcggcgttt
     1801 acgccccccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     1861 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     1921 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     1981 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2041 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2101 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2161 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     2221 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat
     2281 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     2341 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     2401 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     2461 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     2521 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     2581 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     2641 atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttagt
     2701 cgacaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgatgag
     2761 caatgctttt ttataatgcc aactttgtat aataaagttg aacgagaaac gtaaaatgat
     2821 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     2881 acacaacata tccagtcact atgaatcaac tacttagatg gtattagtga cctgtactgc
     2941 agggcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     3001 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     3061 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     3121 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     3181 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     3241 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     3301 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     3361 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     3421 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     3481 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     3541 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     3601 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     3661 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     3721 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     3781 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     3841 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     3901 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     3961 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     4021 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4081 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4141 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4201 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4261 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4321 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4381 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4441 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4501 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4561 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4621 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4681 ggttcctggc cttttgctgg ccttttgctc acatgtt
//