pENTR4

LOCUS       Exported                2720 bp ds-DNA     circular SYN 13-AUG-2016
DEFINITION  synthetic circular DNA
KEYWORDS    pENTR4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2720)
  AUTHORS   Biofeng Lab.
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..2720
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL1"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             615..920
                     /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(949..1048)
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1171..1980
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2070..2658
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac caattcagtc
      481 gactggatcc ggtaccgaat tcgcttacta aaagccagat aacagtatgc gtatttgcgc
      541 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
      601 ggtgtgcttc tagaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
      661 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
      721 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
      781 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
      841 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
      901 tgatgttctg gggaatatag aattcgcggc cgcactcgag atatctagac ccagctttct
      961 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     1021 ctatcagtca aaataaaatc attatttgcc atccagctgc agctctggcc cgtgtctcaa
     1081 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1141 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1201 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1261 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     1321 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     1381 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctggt
     1441 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     1501 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1561 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     1621 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1681 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     1741 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1801 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     1861 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     1921 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     1981 tcagaattgg ttaattggtt gtaacattat tcagattggg ccccgttcca ctgagcgtca
     2041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2701 tggccttttg ctcacatgtt
//