pDONR221-P5P4

LOCUS       Exported                4726 bp ds-DNA     circular SYN 10-AUG-2016
DEFINITION  synthetic circular DNA
KEYWORDS    pDONR221-P5P4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4726)
  AUTHORS   Biofeng Lab.
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4726
                     /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    570..801
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP5"
                     /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    complement(2705..2936)
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP4"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2968..2986)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2991..3007)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3120..3929
                     /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      4076..4664
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa
      721 aaaacagact acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt
      781 agatggtatt agtgacctgt agtcgaccga 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 cgactacagg tcactaatac catctaagta gttgattcat agtgactgga tatgttgtgt
     2761 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     2821 atcattttac gtttctcgtt caacttttct atacaaagtt ggcattataa gaaagcattg
     2881 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2941 tccagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg
     3001 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     3061 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     3121 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     3181 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     3241 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     3301 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     3361 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3421 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3481 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3541 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3601 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3661 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3721 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3781 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3841 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3901 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     3961 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     4021 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4081 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4141 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4201 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4261 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4321 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4381 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4441 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4501 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4561 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4621 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4681 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//