LOCUS       Exported                8849 bp ds-DNA     circular SYN 28-NOV-2016
DEFINITION  EZ-Tet-pLKO-Blast.
ACCESSION   .
VERSION     .
KEYWORDS    EZ-Tet-pLKO-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 28, 2016 from SnapGene Viewer 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8849
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    452..547
                     /note="H1/TetO Promoter"
     misc_feature    555..775
                     /note="Stuffer"
     promoter        983..1493
                     /note="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     intron          1502..2074
                     /note="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     CDS             2158..2775
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /note="TetR"
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
     misc_feature    2813..3386
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3404..3802
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /note="BlastR"
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3997..4230
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             5391..6251
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     LTR             7706..7886
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7933..8058
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8551..8784
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
       61 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
      121 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
      181 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
      241 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      301 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
      361 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
      421 atcgatcacg agactagcct cgagcggccg caatatttgc atgtcgctat gtgttctggg
      481 aaatcaccat aaacgtgaaa tccctatcag tgatagagac ttataagttc cctatcagtg
      541 atagagacgc tagcgtgttg taaatgagca cacaaaatac acatgctaaa atattatatt
      601 ctatgacctt tataaaatca accaaaatct tctttttaat aactttagta tcaataatta
      661 gaatttttat gttccttttt gcaaactttt aataaaaatg agcaaaataa aaaaacgcta
      721 gttttagtaa ctcgcgttgt tttcttcacc tttaataata gctactccac cacttgaatt
      781 ctcgacctcg agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
      841 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
      901 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
      961 tccactttgg ccgcggctcg agggggttgg ggttgcgcct tttccaaggc agccctgggt
     1021 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     1081 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     1141 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     1201 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     1261 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     1321 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     1381 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     1441 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc
     1501 tgtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat tcatgttata
     1561 tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct tgtatcacca
     1621 tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac cattgtctcc
     1681 tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc atttgtaacg
     1741 aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct aatcactttt
     1801 ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga gaacaattgt
     1861 tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc gtggaaatat
     1921 tcttattggt agaaacaact acatcctggt catcatcctg cctttctctt tatggttaca
     1981 atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc
     2041 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     2101 gctgtctcat cattttggca aagaattgta atacgactca ctatagggcg aattgatatg
     2161 tctagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga
     2221 atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg
     2281 tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat
     2341 aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat
     2401 aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta
     2461 ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc
     2521 caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact
     2581 ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct
     2641 actactgata gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt
     2701 gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt
     2761 aaatgtgaaa gtgggtccgc gtacagcggc tcccgggagt tctagggatc tgcccctctc
     2821 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     2881 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     2941 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3001 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     3061 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     3121 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     3181 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     3241 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     3301 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3361 ttttcctttg aaaaacacga tgataaggat ccggaactaa accatggcca agcctttgtc
     3421 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     3481 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     3541 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     3601 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     3661 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     3721 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     3781 tggttatgtg tgggagggct aagcacaatg acgtcgaggt gcccgaagga ccgcgcacct
     3841 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
     3901 aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac aaggcagctg
     3961 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     4021 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     4081 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4141 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4201 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     4261 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     4321 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4381 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     4441 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4501 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4561 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     4621 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     4681 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4741 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4801 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4861 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4921 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4981 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5041 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5101 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5161 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5221 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     5281 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5341 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5401 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5461 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5521 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5581 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5641 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5701 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5761 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5821 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5881 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5941 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6001 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6061 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6121 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6181 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6241 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6301 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6361 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6421 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6481 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6541 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6601 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6661 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6721 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6781 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6841 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6901 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6961 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7021 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7081 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7141 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7201 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7261 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7321 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7381 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     7441 ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt
     7501 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
     7561 ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
     7621 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
     7681 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
     7741 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     7801 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     7861 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     7921 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     7981 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     8041 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     8101 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     8161 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     8221 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     8281 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     8341 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     8401 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     8461 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     8521 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     8581 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     8641 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     8701 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     8761 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     8821 cactgctgtg ccttggaatg ctagttgga
//
