LOCUS       Exported                4718 bp ds-DNA     circular SYN 09-AUG-2016
DEFINITION  synthetic circular DNA
KEYWORDS    pDONR222
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4718)
  AUTHORS   Biofeng Lab.
FEATURES             Location/Qualifiers
     source          1..4718
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      58..85
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      177..263
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     327..343
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    360..591
                     /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(987..1292)
                     /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(1642..2295)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     promoter        complement(2296..2398)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2543..2774)
                     /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        complement(2793..2811)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2816..2832)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2899..3714)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        3722..3777
                     /gene="tetR"
                     /note="tetR/tetA promoters"
                     /note="overlapping promoters for bacterial tetR and tetA"
     rep_origin      complement(4104..4692)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtctgacgct cagggaacga cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
       61 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
      121 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
      181 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
      241 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
      301 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
      361 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
      421 cttttttata atgccaactt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
      481 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
      541 acatatccag tcactatgaa tcaactactt agatggtatt agtgacctgt agtcgaccga
      601 cagccttcca aatgttcttc gggtgatgct gccaacttag tcgaccgaca gccttccaaa
      661 tgttcttctc aaacggaatc gtcgtatcca gcctactcgc tattgtcctc aatgccgtat
      721 taaatcataa aaagaaataa ggaaaagagg tgcgagcctc ttttttgtgt gacaaaataa
      781 aaacatctac ctattcatat acgctagtgt catagtcctg aaaatcatct gcatcaagaa
      841 caatttcaca actcttatac ttttctctta caagtcgttc ggcttcatct ggattttcag
      901 cctctatact tactaaacgt gataaagttt ctgtaatttc tactgtatcg acctgcagac
      961 tggctgtgta taagggagcc tgacatttat attccccaga acatcaggtt aatggcgttt
     1021 ttgatgtcat tttcgcggtg gctgagatca gccacttctt ccccgataac ggagaccggc
     1081 acactggcca tatcggtggt catcatgcgc cagctttcat ccccgatatg caccaccggg
     1141 taaagttcac gggagacttt atctgacagc agacgtgcac tggccagggg gatcaccatc
     1201 cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca caaacagacg ataacggctc
     1261 tctcttttat aggtgtaaac cttaaactgc atttcaccag cccctgttct cgtcagcaaa
     1321 agagccgttc atttcaataa accgggcgac ctcagccatc ccttcctgat tttccgcttt
     1381 ccagcgttcg gcacgcagac gacgggcttc attctgcatg gttgtgctta ccagaccgga
     1441 gatattgaca tcatatatgc cttgagcaac tgatagctgt cgctgtcaac tgtcactgta
     1501 atacgctgct tcatagcata cctctttttg acatacttcg ggtatacata tcagtatata
     1561 ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg ttatctggct tttagtaagc
     1621 cggatccacg cggcgtttac gccccccctg ccactcatcg cagtactgtt gtaattcatt
     1681 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     1741 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     1801 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     1861 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     1921 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     1981 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     2041 ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag
     2101 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     2161 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     2221 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     2281 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     2341 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
     2401 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
     2461 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg
     2521 tgatgctgcc aacttagtcg actacaggtc actaatacca tctaagtagt tgattcatag
     2581 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     2641 taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt acaaagttgg
     2701 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2761 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2821 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2881 taaaaatata tcatcatgtt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2941 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3001 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3061 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3121 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     3181 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3241 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     3301 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3361 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     3421 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3481 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3541 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     3601 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3661 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcatagatct
     3721 tttctccatc actgataggg agtggtaaaa taactccatc aatgatagag tgtcaacaac
     3781 atgaccaaaa tcccttaacg tgagttacgc gtattaattg cgttgcgctc actgcccgct
     3841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3901 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4261 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4501 acagagttct tgaagtggtg gcctaactac ggttacacta gaagaacagt atttggtatc
     4561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4681 aaaggatctc aagaagatcc tttgatcttt tctacggg
//