LOCUS       Exported                6801 bp ds-DNA     circular SYN 06-JUL-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMuLE_ENTR_12GLI-FLuc_R4-R3
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6801)
  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..6801
                     /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    complement(603..727)
                     /gene="mutant version of attR"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attR4"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    753..793
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     polyA_signal    1032..1166
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     polyA_signal    1260..1394
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    1428..1466
                     /note="3xGLIBS"
                     /note="Identified in JM94 2017-11-21, sequence was given by
                     collaborator, see lab book 17.8.17. "
     misc_feature    1467..1505
                     /note="3xGLIBS"
                     /note="Identified in JM94 2017-11-21, sequence was given by
                     collaborator, see lab book 17.8.17. "
     misc_feature    1506..1544
                     /note="3xGLIBS"
                     /note="Identified in JM94 2017-11-21, sequence was given by
                     collaborator, see lab book 17.8.17. "
     promoter        1635..1697
                     /note="Mini-TK promoter"
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     misc_feature    1753..3406
                     /note="Luciferase, manually added, JM94 2017-11-21"
     misc_feature    1753..1764
                     /note="Start of Luc"
                     /note="2017-11-21 JM94"
     misc_feature    2576
                     /note="Replaced ""R"" with G"
                     /note=" as in Luc-F sequencing; jm94 2017-11-21"
     misc_feature    2683
                     /note="Replaced ""R"" with G"
                     /note="as in Luc-F sequencing, JM94 2017-11-21"
     misc_feature    3370..3406
                     /note="End of Luc"
     intron          3538..3603
                     /note="small t intron"
                     /note="simian virus 40 (SV40) small t antigen intron"
     CDS             3733..3753
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     polyA_signal    4178..4312
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     protein_bind    4864..4987
                     /gene="mutant version of attR"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attR3"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(5043..5061)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5066..5082)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5195..6004
                     /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      6151..6739
                     /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 tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgggag acccaagctt ggtaccgagc tctatacact ccgctatcgc tacgtgactg
      781 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      901 gttttcaccg tcatcaccga aacgcgcgag gcaggatcag ccatatcaca tttgtagagg
      961 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     1021 caattgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1081 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1141 tcaatgtatc ttatcatgtc tggatcagcc atatcacatt tgtagaggtt ttacttgctt
     1201 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgtta
     1261 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1381 atcatgtctg gatccaagct cagatccaag cttatcgata ccgtcgagtg ggtggtctgg
     1441 gtggtctggg tggtctgggt ggtcgagtgg gtggtctggg tggtctgggt ggtctgggtg
     1501 gtcgagtggg tggtctgggt ggtctgggtg gtctgggtgg tcgacctcga gatccggcaa
     1561 accccgccca gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc ggggcggcgc
     1621 ggtccgaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc
     1681 tgcagcgacc cgcttaacag cgtcaacagc gtgccgcaga tctgagcttg gcattccggt
     1741 actgttggta aaatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatcct
     1801 ctagaggatg gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt
     1861 cctggaacaa ttgcttttac agatgcacat atcgaggtga acatcacgta cgcggaatac
     1921 ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac
     1981 agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta
     2041 tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac
     2101 agtatgaaca tttcgcagcc taccgtagtg tttgtttcca aaaaggggtt gcaaaaaatt
     2161 ttgaacgtgc aaaaaaaatt accaataatc cagaaaatta ttatcatgga ttctaaaacg
     2221 gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt
     2281 aatgaatacg attttgtacc agagtccttt gatcgtgaca aaacaattgc actgataatg
     2341 aattcctctg gatctactgg gttacctaag ggtgtggccc ttccgcatag aactgcctgc
     2401 gtcagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg
     2461 attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg
     2521 atatgtggat ttcgagtcgt cttaatgtat agatttgaag aaaagctgtt tttacgatcc
     2581 cttcaggatt acaaaattca aagtgcgttg ctagtaccaa ccctattttc attcttcgcc
     2641 aaaagcactc tgattganaa atacgattta tctaatttac acgaaattgc ttctgggggg
     2701 cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat
     2761 acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     2821 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     2881 ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca gaggacctat
     2941 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     3001 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatagttga
     3061 ccgcttgaag tctttaatta aatacaaagg atatcaggtg gcccccgctg aattggaatc
     3121 gatattgtta caacacccca acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga
     3181 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     3241 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     3301 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     3361 agagatcctc ataaaggcca agaagggcgg aaagtccaaa ttgtaaaatg taactgtatt
     3421 cagcgatgac gaaattctta gctattgtaa tgactctaga ggatctttgt gaaggaacct
     3481 tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta
     3541 aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt
     3601 tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa
     3661 aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa
     3721 cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa
     3781 ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt
     3841 tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta
     3901 acctttataa gtaggcataa cagttataat cataacatac tgttttttct tactccacac
     3961 aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta
     4021 atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat
     4081 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4141 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4201 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4261 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcgatcccca
     4321 ggaagctcct ctgtgtcctc ataaacccta acctcctcta cttgagagga cattccaatc
     4381 ataggctgcc catccaccct ctgtgtcctc ctgttaatta ggtcacttaa caaaaaggaa
     4441 attgggtagg ggtttttcac agaccgcttt ctaagggtaa ttttaaaata tctgggaagt
     4501 cccttccact gctgtgttcc agaagtgttg gtaaacagcc cacaaatgtc aacagcagaa
     4561 acatacaagc tgtcagcttt gcacaagggc ccaacaccct gctcatcaag aagcactgtg
     4621 gttgctgtgt tagtaatgtg caaaacagga ggcacatttt ccccacctgt gtaggttcca
     4681 aaatatctag tgttttcatt tttacttgga tcaggaaccc agcactccac tggataagca
     4741 ttatccttat ccaaaacagc cttgtggtca gtgttcatct gctgactagt aacggccgcc
     4801 agtgtgctgg aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga
     4861 ggacaacttt gtataataaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat
     4921 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     4981 cactatgaat caactactta gatggtatta gtgacctgta ctgcagggcg gccgcgatat
     5041 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     5101 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     5161 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     5221 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     5281 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     5341 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     5401 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     5461 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     5521 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     5581 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     5641 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     5701 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     5761 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     5821 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5881 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5941 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     6001 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     6061 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     6121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6301 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6781 ctggcctttt gctcacatgt t
//
