LOCUS       Exported                9194 bp ds-DNA     circular SYN 09-MAY-2017
DEFINITION  Expression Clone, insert flanked by attB1, attB2 sites.
ACCESSION   .
VERSION     .
KEYWORDS    pLenti-Tet-MKK6(DD)-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9194)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, May 9, 2017 from SnapGene Viewer 3.3.4
            http://www.snapgene.com
COMMENT     Entry Clone:   pENTR3C-MKK6-DD, 3484 bps DNA circular (attL1, attL2 
            sites)
            Destination Vector:   pLenti-CMV-TetON-Puro-DEST, 9594 bps DNA 
            circular (attR1, attR2 sites)
            Recombination using LR clonase to produce Expression Clone
FEATURES             Location/Qualifiers
     source          1..9194
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    132..992
                     /label=AmpR
     primer_bind     2137..2153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             2448..2628
                     /label=5' LTR
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2672..2797
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3290..3523
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4012..4129
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4152..4659
                     /label=CMV
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    4661..4700
                     /label=TetO(2x)
     protein_bind    4741..4765
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4800..4829
                     /label=Myc-Tag
     misc_feature    4833..5837
                     /label=MKK6-DD
     misc_feature    5451..5465
                     /label=S207D, T211D
     protein_bind    complement(6021..6045)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6076..6664
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     promoter        6678..7177
                     /label=PGK
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             7198..7797
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             7935..8168
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8576..8592)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt
     2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga
     2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
     3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag
     4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgataagct
     4141 tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4201 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4261 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4321 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4381 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4441 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4501 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4561 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4621 ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag tgatagagat
     4681 ctccctatca gtgatagaga tcgtcgacta gtccagtgtg gtggaattct gcagatatca
     4741 acaagtttgt acaaaaaagc aggctcttta aaggaaccaa ttcagtcgac accatgccgg
     4801 agcagaagct gatatccgag gaggacctgg ccatggctca gtcgaaaggc aagaagcgaa
     4861 accctggcct taaaattcca aaagaagcat ttgaacaacc tcagaccagt tccacaccac
     4921 ctcgagattt agactccaag gcttgcattt ctattggaaa tcagaacttt gaggtgaagg
     4981 cagatgacct ggagcctata atggaactgg gacgaggtgc gtacggggtg gtggagaaga
     5041 tgcggcacgt gcccagcggg cagatcatgg cagtgaagcg gatccgagcc acagtaaata
     5101 gccaggaaca gaaacggcta ctgatggatt tggatatttc catgaggacg gtggactgtc
     5161 cattcactgt caccttttat ggcgcactgt ttcgggaggg tgatgtgtgg atctgcatgg
     5221 agctcatgga tacatcacta gataaattct acaaacaagt tattgataaa ggccagacaa
     5281 ttccagagga catcttaggg aaaatagcag tttctattgt aaaagcatta gaacatttac
     5341 atagtaagct gtctgtcatt cacagagacg tcaagccttc taatgtactc atcaatgctc
     5401 tcggtcaagt gaagatgtgc gattttggaa tcagtggcta cttggtggac gatgttgcta
     5461 aagatattga tgcaggttgc aaaccataca tggcccctga aagaataaac ccagagctca
     5521 accagaaggg atacagtgtg aagtctgaca tttggagtct gggcatcacg atgattgagt
     5581 tggccatcct tcgatttccc tatgattcat ggggaactcc atttcagcag ctcaaacagg
     5641 tggtagagga gccatcgcca caactcccag cagacaagtt ctctgcagag tttgttgact
     5701 ttacctcaca gtgcttaaag aagaattcca aagaacggcc tacataccca gagctaatgc
     5761 aacatccatt tttcacccta catgaatcca aaggaacaga tgtggcatct tttgtaaaac
     5821 tgattcttgg agactaactc gagatcgatt agactagtct agaaattcac cccaccagtg
     5881 caggctgcct atcagaaagt ggtggctggt gtggctaatg ccctggccca caagtatcac
     5941 taagctcgct ttcttgctgt ccaatttcta ttaaaggttc ctttgttccc taagtccgcg
     6001 gccgcactcg agatatctag acccagcttt cttgtacaaa gtggttgata tccagcacag
     6061 tggcggccgc tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6121 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6181 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6241 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6301 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6361 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6421 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     6481 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6541 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6601 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6661 ccgcctggaa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     6721 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     6781 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     6841 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     6901 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     6961 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     7021 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     7081 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     7141 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg
     7201 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     7261 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     7321 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     7381 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     7441 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     7501 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     7561 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     7621 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     7681 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     7741 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc
     7801 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atctcgaggg
     7861 cccggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     7921 agaaaagggg ggactggaag ggctagctca ctcccaacga agacaagatc tgctttttgc
     7981 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctgcctagg
     8041 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     8101 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     8161 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     8221 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     8281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     8341 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     8401 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     8461 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     8521 ttttgcgtcg agacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     8581 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     8641 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     8701 cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa
     8761 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     8821 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     8881 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     8941 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     9001 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     9061 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     9121 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     9181 cgtttacaat ttcc
//
