LOCUS       V010423                 9550 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010423
VERSION     V010423
KEYWORDS    pLVX-rHom-Sec1.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9550)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9550)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1721..1765
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2027..2144
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2201..2504
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    2804..2815
                     /label="5' MCS"
                     /note="5' MCS"
                     /note="multiple cloning site upstream of DmrD"
     sig_peptide     2822..2899
                     /label="hGH signal sequence"
                     /note="human growth hormone signal sequence"
     CDS             2906..3226
                     /label="DmrD"
                     /note="dimeric F36M mutant of FK506-binding protein FKBP12"
     CDS             3233..3553
                     /codon_start=1
                     /product="dimeric F36M mutant of FK506-binding protein
                     FKBP12"
                     /label="dimeric F36M mutant of FK506-binding protein FK"
                     /note="DmrD"
                     /note="dimerization can be reversed by ligand binding"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3560..3880
                     /codon_start=1
                     /product="dimeric F36M mutant of FK506-binding protein
                     FKBP12"
                     /label="dimeric F36M mutant of FK506-binding protein FK"
                     /note="DmrD"
                     /note="dimerization can be reversed by ligand binding"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3887..4207
                     /codon_start=1
                     /product="dimeric F36M mutant of FK506-binding protein
                     FKBP12"
                     /label="dimeric F36M mutant of FK506-binding protein FK"
                     /note="DmrD"
                     /note="dimerization can be reversed by ligand binding"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     misc_feature    4227..4240
                     /label="3' MCS"
                     /note="3' MCS"
                     /note="multiple cloning site downstream of DmrD"
     misc_feature    4253..4839
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4859..5455
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5472..6060
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6268..6901
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7030..7046)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7054..7070)
                     /label="lac operator"
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7078..7108)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7123..7144)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7432..8017)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8191..9048)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9049..9153)
                     /label="AmpR promoter"
     polyA_signal    9201..9335
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation 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 actacaaaaa 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 cgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacaccaa 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 tagctagagg atctatttcc ggtgaattcc tcgagactag
     2821 tatggctaca ggctcccgga cgtccctgct cctggctttt ggcctgctct gcctgccctg
     2881 gcttcaagag ggcagtgcct ctagaggagt gcaggtggag actatctccc caggagacgg
     2941 gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga
     3001 tggaaagaaa atggattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa
     3061 gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc
     3121 caaactgact atatctccag attatgccta tggtgccact gggcacccag gcatcatccc
     3181 accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaaacta gaggagtgca
     3241 ggtggagact atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt
     3301 ggtgcactac accgggatgc ttgaagatgg aaagaaaatg gattcctccc gggacagaaa
     3361 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
     3421 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
     3481 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
     3541 tctaaaactg gaaactagag gagtgcaggt ggagactatc tccccaggag acgggcgcac
     3601 cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg aagatggaaa
     3661 gaaaatggat tcctcccggg acagaaacaa gccctttaag tttatgctag gcaagcagga
     3721 ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcaga gagccaaact
     3781 gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca tcccaccaca
     3841 tgccactctc gtcttcgatg tggagcttct aaaactggaa actagaggag tgcaggtgga
     3901 gactatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca
     3961 ctacaccggg atgcttgaag atggaaagaa aatggattcc tcccgggaca gaaacaagcc
     4021 ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca
     4081 gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac
     4141 tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa
     4201 actggaaact agtgctagaa accccagcgg ccgcggatcc tacgtatgac cgcccctctc
     4261 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     4321 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     4381 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     4441 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     4501 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4561 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     4621 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     4681 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     4741 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     4801 ttttcctttg aaaaacacga tgataagctt gccacaaccc acaaggagac gaccttccat
     4861 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     4921 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     4981 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     5041 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     5101 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     5161 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     5221 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     5281 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     5341 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     5401 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctagac
     5461 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5521 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5581 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5641 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5701 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5761 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5821 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     5881 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5941 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6001 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6061 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     6121 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     6181 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     6241 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     6301 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     6361 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     6421 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     6481 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     6541 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     6601 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     6661 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     6721 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6781 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6841 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6901 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6961 gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct agagcttggc
     7021 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7081 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7141 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7201 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7261 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7321 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7861 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     7981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8101 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8161 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8221 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8281 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8341 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8401 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8461 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8521 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8581 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8641 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8701 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8761 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8821 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8881 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8941 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9001 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9061 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9181 cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca
     9241 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     9301 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     9361 atcatcccaa acttcccacc ccatacccta ttaccactgc caattaccta gtggtttcat
     9421 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
     9481 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa
     9541 acttagtagt
//
