LOCUS       V006704                 8798 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V006704
VERSION     V006704
KEYWORDS    pLenti CMV GFP Neo (657-2)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8798)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO,
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
   PUBMED   19657394
REFERENCE   2  (bases 1 to 8798)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8798)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        58..162
                     /label="AmpR promoter"
     CDS             163..1020
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1194..1782
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     1936..1953
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    2070..2091
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2106..2136
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2144..2160
                     /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)."
     primer_bind     2168..2184
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2205..2223
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2251..2477
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2478..2658
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2705..2830
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    3323..3556
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             3741..3785
                     /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             3934..3975
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    4052..4168
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        4191..4494
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4495..4698
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     4695..4719
                     /label="LNCX"
                     /note="Human CMV promoter, forward primer"
     CDS             4794..5510
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    5529..6117
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        6160..6659
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             6673..7473
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase from Tn5"
     LTR             7567..7800
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7872..8006
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      8033..8168
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(8189..8207)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8217..8233)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      8375..8798
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
       61 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      121 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      181 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      241 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      301 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      361 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
      421 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      481 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      541 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      601 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      661 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
      721 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      781 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      841 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
      901 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      961 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1021 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1081 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1141 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1201 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1261 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1321 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1381 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1441 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1501 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1561 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1621 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1681 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1741 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1801 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1861 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     1921 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     1981 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2041 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     2101 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2161 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     2221 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     2281 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     2341 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     2401 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     2461 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2521 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     2581 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     2641 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     2701 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     2761 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     2821 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     2881 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     2941 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     3001 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     3061 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     3121 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     3181 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     3241 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     3301 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     3361 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     3421 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     3481 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     3541 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     3601 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     3661 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     3721 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     3781 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     3841 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     3901 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     3961 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     4021 tagtgaacgg atctcgacgg tatcggttaa cttttaaaag aaaagggggg attggggggt
     4081 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     4141 aaaaacaaat tacaaaattc aaaattttat cgataagctt gggagttccg cgttacataa
     4201 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     4261 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     4321 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     4381 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     4441 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     4501 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     4561 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     4621 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     4681 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
     4741 tgttttgacc tccatagaag acaccgactc tagaggatcc accggtcgcc accatggtga
     4801 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4861 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4921 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4981 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     5041 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     5101 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     5161 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     5221 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     5281 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     5341 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5401 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5461 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     5521 gcgtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     5581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     5641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5701 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     5761 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     5821 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     5881 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     5941 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6001 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6061 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     6121 gaattctgca gatatccgga ctagtgatct aattctaccg ggtaggggag gcgcttttcc
     6181 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     6241 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     6301 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     6361 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     6421 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     6481 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     6541 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     6601 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     6661 cctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg ttctccggcc
     6721 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     6781 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     6841 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     6901 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     6961 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     7021 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     7081 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     7141 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     7201 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     7261 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     7321 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     7381 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     7441 atcgccttct atcgccttct tgacgagttc ttctgagggg atccgtcgag ggcccggtac
     7501 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     7561 ggggactgga agggctagct cactcccaac gaagacaaga tctgcttttt gcttgtactg
     7621 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7681 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7741 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7801 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     7861 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7921 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     7981 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     8041 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     8101 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     8161 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     8221 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     8281 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     8341 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     8401 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     8461 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     8521 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     8581 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     8641 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     8701 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     8761 tggttaaaaa atgagctgat ttaacaaaaa tttaacgc
//
