pDONR221-P1P4

LOCUS       Exported                4774 bp ds-DNA     circular SYN 10-AUG-2016
DEFINITION  synthetic circular DNA
KEYWORDS    pDONR221-P1P4
  AUTHORS   Biofeng Lab.
FEATURES             Location/Qualifiers
     source          1..4774
                     /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="attP1"
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)201 version)"
     CDS             complement(1197..1502)
                     /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(1852..2505)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     promoter        complement(2506..2608)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2753..2984)
                     /gene="mutant version of attP"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attP4"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3016..3034)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3039..3055)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3168..3977
                     /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      4124..4712
                     /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 tgtacaaaaa
      661 agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa
      721 aaaacagact acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt
      781 agatggtatt agtgacctgt agtcgaccga cagccttcca aatgttcttc gggtgatgct
      841 gccaacttag tcgaccgaca gccttccaaa tgttcttctc aaacggaatc gtcgtatcca
      901 gcctactcgc tattgtcctc aatgccgtat taaatcataa aaagaaataa gaaaaagagg
      961 tgcgagcctc ttttttgtgt gacaaaataa aaacatctac ctattcatat acgctagtgt
     1021 catagtcctg aaaatcatct gcatcaagaa caatttcaca actcttatac ttttctctta
     1081 caagtcgttc ggcttcatct ggattttcag cctctatact tactaaacgt gataaagttt
     1141 ctgtaatttc tactgtatcg acctgcagac tggctgtgta taagggagcc tgacatttat
     1201 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca
     1261 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc
     1321 cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc
     1381 agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc
     1441 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc
     1501 atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac
     1561 ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc
     1621 attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac
     1681 tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg
     1741 acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat
     1801 acgcatactg ttatctggct tttagtaagc cggatccacg cggcgtttac gccccccctg
     1861 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     1921 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     1981 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     2041 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     2101 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     2161 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     2221 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     2281 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     2341 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     2401 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     2461 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     2521 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     2581 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg
     2641 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac
     2701 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttagtcg actacaggtc
     2761 actaatacca tctaagtagt tgattcatag tgactggata tgttgtgttt tacagtatta
     2821 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     2881 ttctcgttca acttttctat acaaagttgg cattataaga aagcattgct tatcaatttg
     2941 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagg
     3001 gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     3061 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     3121 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     3181 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     3241 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3301 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3361 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     3421 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     3481 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3541 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3601 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3661 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3721 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3781 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3841 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3901 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3961 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     4021 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     4081 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4141 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4201 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4261 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4321 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4381 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4441 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4501 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4561 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4621 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4681 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4741 tcctggcctt ttgctggcct tttgctcaca tgtt
//