LOCUS       Exported                3693 bp ds-DNA     circular SYN 06-JUL-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMuLE_ENTR_TOP-NLuc1.1_L5-L4
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 3693)
  AUTHORS   Haider Sami
  TITLE     Direct Submission
  JOURNAL   Exported Friday, July 6, 2018 from SnapGene 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3693
                     /organism="recombinant plasmid"
                     /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..665
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL5"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    699..709
                     /note="TCF binding site"
     misc_feature    714..724
                     /note="TCF binding site"
     misc_feature    729..739
                     /note="TCF binding site"
     misc_feature    745..755
                     /note="TCF binding site"
     misc_feature    760..770
                     /note="TCF binding site"
     misc_feature    775..785
                     /note="TCF binding site"
     misc_feature    790..800
                     /note="TCF binding site"
     promoter        840..871
                     /note="minP"
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    899..903
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             933..1448
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /note="Nluc"
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    1488..1709
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1808..1903)
                     /gene="mutant version of attL"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attL4"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1935..1953)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1958..1974)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2087..2896
                     /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      3043..3631
                     /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 agttgggaga cccaagcttg gtaccgagct cttacgcgag atcaaagggg gtaagatcaa
      721 agggggtaag atcaaagggg cgcgagatca aagggggtaa gatcaaaggg ggtaagatca
      781 aagggggtaa gatcaaaggg gcgcgcccgc gtgctagccc gggctcgaga tctagactct
      841 agagggtata taatggaagc tcgaattcca gcttggcatt ccggtactgt tggtaaaaag
      901 cttggcaatc cggtactgtt ggtaaagcca ccatggtctt cacactcgaa gatttcgttg
      961 gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt
     1021 ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg
     1081 gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg
     1141 accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact
     1201 ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg
     1261 actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa
     1321 cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct
     1381 ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc
     1441 tggcgtaagg ccgcgactct agagtcgggg cggccggccg cttcgagcag acatgataag
     1501 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1561 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1621 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     1681 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccactag taacggccgc
     1741 cagtgtgctg gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag
     1801 aggcacccaa cttttctata caaagttggc attataagaa agcattgctt atcaatttgt
     1861 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcaggg
     1921 cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1981 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2041 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2101 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2161 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2221 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2281 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2341 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2401 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2461 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2521 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2581 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2641 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2701 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2761 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2821 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2881 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2941 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3001 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3061 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3121 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3181 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3241 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3301 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3361 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3421 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3481 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3541 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3601 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3661 cctggccttt tgctggcctt ttgctcacat gtt
//
