LOCUS       Exported                8547 bp ds-DNA     circular SYN 05-MAY-2017
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    JB138
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8547)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported 5 May 2017 from SnapGene 3.0.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8547
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     LTR             827..1137
                     /note="5prime LTR"
     misc_recomb     1933..1966
                     /note="F3 site"
     polyA_signal    complement(1983..2204)
                     /note="SV40\polyA"
                     /note="vntifkey=""25"" - label=SV40polyA - 
                     complement(1983..2204)"
     CDS             complement(2395..2809)
                     /note="BirA-mut [*] [3']"
     misc_difference 2809
                     /note="The\decisive\arginine [3']"
                     /note="vntifkey=""85"" - label=Thedecisivearginine - 
                     5011..5013"
     misc_difference 2811..2812
                     /note="The\decisive\arginine [5']"
                     /note="vntifkey=""85"" - label=Thedecisivearginine - 
                     5011..5013"
     CDS             complement(2811)
                     /note="BirA-WT [*] [5']"
     CDS             complement(2812..3159)
                     /note="BirA-WT [*]"
     CDS             complement(3163..3192)
                     /note="c-Myc tag"
     misc_feature    3199..3246
                     /note="Gly/Ser linker 2"
     CDS             complement(3253..3576)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /note="FKBP"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     promoter        complement(3592..3716)
                     /note="Pcmv min"
     misc_binding    3734..3752
                     /note="tetO"
     misc_binding    3776..3794
                     /note="tetO"
     misc_binding    3818..3836
                     /note="tetO"
     misc_binding    3860..3878
                     /note="tetO"
     misc_binding    3902..3920
                     /note="tetO"
     misc_binding    3944..3962
                     /note="tetO"
     misc_binding    3986..4004
                     /note="tetO"
     promoter        4021..4145
                     /note="Pcmv min"
     misc_feature    4169..4192
                     /note="FLAG tag"
     CDS             4169..4192
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /note="FLAG"
                     /translation="DYKDDDDK"
     CDS             4199..4393
                     /note="BirA-mut [*] [3']"
     misc_feature    4406..4450
                     /note="Gly/Ser linker 1"
     CDS             4466..4744
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /note="FRB"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     misc_recomb     4799..4832
                     /note="FRT"
     misc_recomb     4799..4832
                     /note="FRT site"
     misc_feature    4942..4947
                     /note="putative\polyA\site"
                     /note="vntifkey=""21"" - label=putativepolyAsite - 
                     8487..8492"
     LTR             4979..5216
                     /note="3prime LTR"
     primer_bind     complement(5919..5936)
                     /note="M13-fwd"
     CDS             6007..6075
                     /note="LacZ alpha"
     promoter        6445..6473
                     /note="Amp prom"
     CDS             6713..7372
                     /note="AmpR"
     rep_origin      7524..8152
                     /direction=RIGHT
                     /note="ColE1 origin"
     promoter        8458..8487
                     /note="lac"
     misc_binding    8492..8514
                     /note="LacO"
     primer_bind     8520..8540
                     /note="M13-rev"
ORIGIN
        1 caagctagct ttgctcttag gagtttccta atacatccca aactcaaata tataaagcat
       61 ttgacttgtt ctatgcccta gggggcgggg ggaagctaag ccagcttttt ttaacattta
      121 aaatgttaat tccattttaa atgcacagat gtttttattt cataagggtt tcaatgtgca
      181 tgaatgctgc aatattcctg ttaccaaagc tagtataaat aaaaatagat aaacgtggaa
      241 attacttaga gtttctgtca ttaacgtttc cttcctcagt tgacaacata aatgcgctgc
      301 tgagcaagcc agtttgcatc tgtcaggatc aatttcccat tatgccagtc atattaatta
      361 ctagtcaatt agttgatttt tatttttgac atatacatgt gaatgaaaga ccccacctgt
      421 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
      481 agaatagaaa agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
      541 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
      601 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      661 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
      721 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      781 gcttctcgct tctgttcgcg cgcttatgct ccccgagctc aataaaagag cccacaaccc
      841 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
      901 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
      961 agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga
     1021 cccctgccca gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt
     1081 gtctgtccga ttgtctagtg tctatgactg attttatgcg cctgcgtcgg tactagttag
     1141 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttcgga acacccggcc
     1201 gcaaccctgg gagacgtccc agggacttcg ggggccgttt ttgtggcccg acctgagtcc
     1261 taaaatcccg atcgtttagg actctttggt gcacccccct tagaggaggg atatgtggtt
     1321 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     1381 tgggaccgaa gccgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
     1441 ctgactgtgt ttctgtattt gtctgaaaat atgggcccgg gctagcctgt taccactccc
     1501 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1561 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1621 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1681 ttttcacctg gcccgcatgg acacccagac caggtggggt acatcgtgac ctgggaagcc
     1741 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1801 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1861 tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata tgagatccga
     1921 agttcctatt ccgaagttcc tattcttcaa atagtatagg aacttcagat cataatcagc
     1981 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2041 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2101 tacaaataag gcaatagcat cacaaatttc acaaataagg catttttttc actgcattct
     2161 agttttggtt tgtccaaact catcaatgta tcttatcatg tctggatctc aaatccctcg
     2221 gaagctgcgc ctgtcttagg ttggagtgat acatttttat cacttttacc cgtctttgga
     2281 ttaggcagta gctctgacgg ccctcctgtc ttaggttagt gaaaaatgtc actctcttac
     2341 ccgtcattgg ctgtccagct tagctcgcag gggaggtggt ctggatccat gtttctcctg
     2401 ctcgaacagc tccagagcgg ctctcagctc tctgatcagc atggcggcca gggtgttccg
     2461 gtccaggttg atgccagcct cctgcagggt gatccagccc tggttcacca cgctctcctc
     2521 cactcttctc atggccatgt tgattccggc tccgatcacg atctgagcgg catcgcctgt
     2581 cttgcctgtc agctccacca ggatgccagc cagcttgcgg tcctggaggt acaggtcatt
     2641 aggccacttc actctcacct tgtcggctcc cagctttctc agcacctcgg ccatcacgat
     2701 tccgatcacc agactcagtc cgatggcagc agcaggtccc tgctccagtc tccagaacat
     2761 ggacaggtac aggttggctc cgaatgggct gaaccacttc cgtcctctgc ctcctctgcc
     2821 agcctgctgg tactcggcga tgcaggcgtc gccggacttc agctctccga ttctgtccag
     2881 caggtactga ttggtggagt cgatcacagg cagcacggcc acgcttccgc catccagctg
     2941 tcccaggatc tgcttggcgt tcagcagctg gataggctca ggcaggctgt agcccttgcc
     3001 aggcacggtg aacacgtcca ctccccagtc tctcagtgtc tggatgtgct tgttgatggc
     3061 ggctctgctc attcccaggg tctctcccag ctgctcgcca gagtggaact cgccgttggc
     3121 cagcagggcg atcagcttca ggggcacggt gttgtccttg tcgagatcct cttctgagat
     3181 gagtttttgt tcatgtttgc cccgggaggc gtaggagatc tggccgccgc cggaggagcc
     3241 gccgccacgc gtttccagtt ttagaagctc cacatcgaag acgagagtgg catgtggtgg
     3301 gatgatgcct gggtgcccag tggcaccata ggcataatct ggagatatag tcagtttggc
     3361 tctctgaccc acactcatct gggcaacccc ttcttcccag cctcggatca cctcctgctt
     3421 gcctagcata aacttaaagg gcttgtttct gtcccgggag gaatcaaatt tctttccatc
     3481 ttcaagcatc ccggtgtagt gcaccacgca ggtctggccg cgcttgggga aggtgcgccc
     3541 gtctcctggg gagatggttt ccacctgcac tcccatatcg attagctcga ccgcggaggc
     3601 tggatcggtc ccggtgtctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga
     3661 tctgacggtt cactaaacga gctctgctta tataggcctc ccaccgtaca cgcctacaag
     3721 cttctttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct
     3781 atcactgata gggagtggta aactcgactt tcacttttct ctatcactga tagggagtgg
     3841 taaactcgac tttcactttt ctctatcact gatagggagt ggtaaactcg actttcactt
     3901 ttctctatca ctgataggga gtggtaaact cgactttcac ttttctctat cactgatagg
     3961 gagtggtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgaggg
     4021 taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     4081 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     4141 ccgcggaatt cggccggcct caattatgga ctacaaggac gacgatgaca agattaacgg
     4201 actggctcct tacctgagca gatgggagaa gctggacaac ttcatcaaca gacctgtgaa
     4261 gctgatcatc ggcgacaagg aaatcttcgg catctccaga ggaatcgaca agcagggagc
     4321 tctgctgctg gagcaggacg gaatcatcaa gccctggatg ggcggagaaa tctccctgag
     4381 aagcgcagag aagctcgagg cgaaccagat ctcctacgcc tcccggggcg gcggctcctc
     4441 cggcggcggc gtttgtttaa acatgatcct ctggcatgag atgtggcatg aaggcctgga
     4501 agaggcatct cgtttgtact ttggggaaag gaacgtgaaa ggcatgtttg aggtgctgga
     4561 gcccttgcat gctatgatgg aacggggccc ccagactctg aaggaaacat cctttaatca
     4621 ggcctatggt cgagatttaa tggaggccca agagtggtgc aggaagtaca tgaaatcagg
     4681 gaatgtcaag gacctcaccc aagcctggga cctctattat catgtgttcc gacgaatctc
     4741 aaagcagtta attaaatagc gattaataac gaggatctgg ccgcgaagtt cctattccga
     4801 agttcctatt ctctagaaag tataggaact tcagatccgg attagtccaa tttgttaaag
     4861 acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta
     4921 tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     4981 tgaaagaccc cacctgtagg tttggcaagc tagctcgact tctgttcgcg cgcttctgct
     5041 ccccgagctc gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     5101 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     5161 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacaca
     5221 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat
     5281 agtacttaaa gttacattgg cttccttgaa ataaacatgg agtattcaga atgtgtcata
     5341 aatatttcta attttaagat agtatctcca ttggctttct actttttctt ttattttttt
     5401 ttgtcctctg tcttccattt gttgttgttg ttgtttgttt gtttgtttgt tggttggttg
     5461 gttaattttt ttttaaagat cctacactat agttcaagct agactattag ctactctgta
     5521 acccagggtg accttgaagt catgggtagc ctgctgtttt agccttccca catctaagat
     5581 tacaggtatg agctatcatt tttggtatat tgattgattg attgattgat gtgtgtgtgt
     5641 gtgattgtgt ttgtgtgtgt gactgtgaaa atgtgtgtat gggtgtgtgt gaatgtgtgt
     5701 atgtatgtgt gtgtgtgagt gtgtgtgtgt gtgtgtgcat gtgtgtgtgt gtgactgtgt
     5761 ctatgtgtat gactgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt
     5821 tgtgaaaaaa tattctatgg tagtgagagc caacgctccg gctcaggtgt caggttggtt
     5881 tttgagacag agtctttcac ttagcttgga attaattcac tggccgtcgt tttacaacgt
     5941 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     6001 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     6061 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6121 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     6181 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     6241 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     6301 ccgaaacgcg cgatgacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6361 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6421 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6481 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6541 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6601 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6661 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6721 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6781 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6841 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6901 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6961 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7021 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7081 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7141 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7201 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7261 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7321 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7381 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7441 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7681 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7921 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8221 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     8281 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     8341 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     8401 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     8461 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     8521 aggaaacagc tatgaccatg attacgc
//
