LOCUS       pLVX-Puro-BTK(human)   10047 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019551
VERSION     V019551
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2827..4803
                     /label="BTK(NM_000061)"
                     /note="BTK(NM_000061)"
                     /gene="BTK"
     promoter        4835..5334
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5355..5954
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5968..6556
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6763..7396
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7525..7541)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7549..7565
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7573..7603)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7618..7639
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7927..8515)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8686..9546)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9547..9651)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9699..9833
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 gccaccatgg ccgcagtgat tctggagagc atctttctga agcgatccca acagaaaaag
     2881 aaaacatcac ctctaaactt caagaagcgc ctgtttctct tgaccgtgca caaactctcc
     2941 tactatgagt atgactttga acgtgggaga agaggcagta agaagggttc aatagatgtt
     3001 gagaagatca cttgtgttga aacagtggtt cctgaaaaaa atcctcctcc agaaagacag
     3061 attccgagaa gaggtgaaga gtccagtgaa atggagcaaa tttcaatcat tgaaaggttc
     3121 ccttatccct tccaggttgt atatgatgaa gggcctctct acgtcttctc cccaactgaa
     3181 gaactaagga agcggtggat tcaccagctc aaaaacgtaa tccggtacaa cagtgatctg
     3241 gttcagaaat atcacccttg cttctggatc gatgggcagt atctctgctg ctctcagaca
     3301 gccaaaaatg ctatgggctg ccaaattttg gagaacagga atggaagctt aaaacctggg
     3361 agttctcacc ggaagacaaa aaagcctctt cccccaacgc ctgaggagga ccagatcttg
     3421 aaaaagccac taccgcctga gccagcagca gcaccagtct ccacaagtga gctgaaaaag
     3481 gttgtggccc tttatgatta catgccaatg aatgcaaatg atctacagct gcggaagggt
     3541 gatgaatatt ttatcttgga ggaaagcaac ttaccatggt ggagagcacg agataaaaat
     3601 gggcaggaag gctacattcc tagtaactat gtcactgaag cagaagactc catagaaatg
     3661 tatgagtggt attccaaaca catgactcgg agtcaggctg agcaactgct aaagcaagag
     3721 gggaaagaag gaggtttcat tgtcagagac tccagcaaag ctggcaaata tacagtgtct
     3781 gtgtttgcta aatccacagg ggaccctcaa ggggtgatac gtcattatgt tgtgtgttcc
     3841 acacctcaga gccagtatta cctggctgag aagcaccttt tcagcaccat ccctgagctc
     3901 attaactacc atcagcacaa ctctgcagga ctcatatcca ggctcaaata tccagtgtct
     3961 caacaaaaca agaatgcacc ttccactgca ggcctgggat acggatcatg ggaaattgat
     4021 ccaaaggacc tgaccttctt gaaggagctg gggactggac aatttggggt agtgaagtat
     4081 gggaaatgga gaggccagta cgacgtggcc atcaagatga tcaaagaagg ctccatgtct
     4141 gaagatgaat tcattgaaga agccaaagtc atgatgaatc tttcccatga gaagctggtg
     4201 cagttgtatg gcgtctgcac caagcagcgc cccatcttca tcatcactga gtacatggcc
     4261 aatggctgcc tcctgaacta cctgagggag atgcgccacc gcttccagac tcagcagctg
     4321 ctagagatgt gcaaggatgt ctgtgaagcc atggaatacc tggagtcaaa gcagttcctt
     4381 caccgagacc tggcagctcg aaactgtttg gtaaacgatc aaggagttgt taaagtatct
     4441 gatttcggcc tgtccaggta tgtcctggat gatgaataca caagctcagt aggctccaaa
     4501 tttccagtcc ggtggtcccc accggaagtc ctgatgtata gcaagttcag cagcaaatct
     4561 gacatttggg cttttggggt tttgatgtgg gaaatttact ccctggggaa gatgccatat
     4621 gagagattta ctaacagtga gactgctgaa cacattgccc aaggcctacg tctctacagg
     4681 cctcatctgg cttcagagaa ggtatatacc atcatgtaca gttgctggca tgagaaagca
     4741 gatgagcgtc ccactttcaa aattcttctg agcaatattc tagatgtcat ggatgaagaa
     4801 tcctgaggat cccgcgactc tagataattc taccgggtag gggaggcgct tttcccaagg
     4861 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     4921 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     4981 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     5041 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     5101 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     5161 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     5221 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
     5281 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc
     5341 agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     5401 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5461 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5521 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5581 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5641 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5701 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5761 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5821 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5881 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5941 agcccggtgc ctgaccgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
     6001 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6061 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6121 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6181 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6241 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6301 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6361 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6421 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6481 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6541 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa
     6601 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     6661 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     6721 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
     6781 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     6841 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     6901 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     6961 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     7021 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     7081 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     7141 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     7201 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     7261 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7321 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7381 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     7441 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac cgtcgacctc
     7501 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     7561 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7621 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7681 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7741 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7801 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7861 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     8041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     8101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     8161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     8221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     8281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     8401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8641 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8701 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8761 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8821 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8881 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8941 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     9001 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     9061 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     9121 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9181 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9241 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9301 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9361 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9421 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9481 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9541 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9601 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9661 ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata
     9721 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     9781 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
     9841 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
     9901 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa
     9961 gaaattgtat ttgttaaata tgtactacaa acttagtagt ttttaaagaa attgtatttg
    10021 ttaaatatgt actacaaact tagtagt
//
