LOCUS       V010424                 9160 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010424
VERSION     V010424
KEYWORDS    pLVX-rHom-Nuc1.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9160)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9160)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9160
                     /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"
     CDS             2822..2824
                     /codon_start=1
                     /product="start codon"
                     /label="start codon"
                     /note="ATG"
                     /translation="M"
     CDS             2825..2845
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2852..3172
                     /label="DmrD"
                     /note="dimeric F36M mutant of FK506-binding protein FKBP12"
     CDS             3179..3499
                     /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             3506..3826
                     /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    3837..3850
                     /label="3' MCS"
                     /note="3' MCS"
                     /note="multiple cloning site downstream of DmrD"
     misc_feature    3863..4449
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4469..5065
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5082..5670
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5878..6511
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6640..6656)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6664..6680)
                     /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(6688..6718)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6733..6754)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7042..7627)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7801..8658)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8659..8763)
                     /label="AmpR promoter"
     polyA_signal    8811..8945
                     /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 tatgcctaag aagaagagaa aggtgtctag aggagtgcag gtggagacta tctccccagg
     2881 agacgggcgc accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct
     2941 tgaagatgga aagaaaatgg attcctcccg ggacagaaac aagcccttta agtttatgct
     3001 aggcaagcag gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca
     3061 gagagccaaa ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat
     3121 catcccacca catgccactc tcgtcttcga tgtggagctt ctaaaactgg aaactagagg
     3181 agtgcaggtg gagactatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac
     3241 ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaaatggatt cctcccggga
     3301 cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga
     3361 aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc
     3421 ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt
     3481 ggagcttcta aaactggaaa ctagaggagt gcaggtggag actatctccc caggagacgg
     3541 gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga
     3601 tggaaagaaa atggattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa
     3661 gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc
     3721 caaactgact atatctccag attatgccta tggtgccact gggcacccag gcatcatccc
     3781 accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaaacta gttatagcgg
     3841 ccgcggatcc tacgtatgac cgcccctctc cctccccccc ccctaacgtt actggccgaa
     3901 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     3961 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     4021 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     4081 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     4141 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     4201 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     4261 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     4321 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     4381 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt
     4441 gccacaaccc acaaggagac gaccttccat gaccgagtac aagcccacgg tgcgcctcgc
     4501 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4561 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4621 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4681 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4741 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4801 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4861 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4921 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4981 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5041 gtgcatgacc cgcaagcccg gtgcctagac gcgtctggaa caatcaacct ctggattaca
     5101 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5161 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5221 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5281 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5341 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5401 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5461 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     5521 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5581 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5641 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga
     5701 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
     5761 agaagcacaa gaggaggagg aggtgggttt ttccagtcac acctcaggta cctttaagac
     5821 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg
     5881 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     5941 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
     6001 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
     6061 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     6121 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
     6181 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
     6241 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
     6301 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
     6361 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6421 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6481 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6541 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt
     6601 tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6781 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6841 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7441 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7561 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7621 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7681 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7741 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7801 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7861 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7921 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7981 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8041 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8101 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8161 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8221 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8281 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8341 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8401 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8461 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8521 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8581 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8641 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8701 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8761 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta
     8821 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8881 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     8941 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta
     9001 ttaccactgc caattaccta gtggtttcat ttactctaaa cctgtgattc ctctgaatta
     9061 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa
     9121 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
