LOCUS       pLenti_CMV/TO_eG        8771 bp DNA     circular SYN 13-MAY-2021
DEFINITION  3rd gen lentiviral eGFP expression vector, CMV/TO promoter, Puro .
ACCESSION   .
VERSION     .
KEYWORDS    pLenti CMV/TO eGFP Puro (w159-1).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8771)
  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 8771)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8771)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /label=AmpR promoter
     CDS             132..989
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1163..1751
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1905..1922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2039..2060
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2075..2105
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /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     2137..2153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2174..2192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2220..2447
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2448..2628
                     /label=5' LTR (truncated)
                     /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."
     CDS             3707..3751
                     /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             3900..3941
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    4012..4129
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        4152..4455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4456..4659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     4656..4680
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    4764..4788
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4854..5570
                     /label=EGFP
                     /note="enhanced GFP"
     protein_bind    complement(5598..5622)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5653..6241
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        6255..6754
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             6775..7371
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     LTR             7512..7745
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7817..7951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      7957..8092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(8125..8143)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8153..8169)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      8314..8769
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
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 gtttagtgaa ccgtcagatc
     4681 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact ctagtccagt
     4741 gtggtggaat tctgcagata tcaacaagtt tgtacaaaaa agcaggctcc accatgggaa
     4801 ccaattcagt cgactggtac cgcgggcccg ggatccaccg gccggtcgcc accatggtga
     4861 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4921 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4981 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     5041 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     5101 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     5161 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     5221 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     5281 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     5341 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     5401 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5461 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5521 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     5581 gcactcgaga tatctagacc cagctttctt gtacaaagtg gttgatatcc agcacagtgg
     5641 cggccgctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5701 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5761 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5821 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5881 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5941 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6001 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     6061 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6121 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6181 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6241 cctggaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     6301 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     6361 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     6421 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     6481 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     6541 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     6601 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     6661 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     6721 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc
     6781 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     6841 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     6901 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     6961 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     7021 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7081 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     7141 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7201 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     7261 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     7321 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     7381 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc tcgagggccc
     7441 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     7501 aaagggggga ctggaagggc tagctcactc ccaacgaaga caagatctgc tttttgcttg
     7561 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct gcctagggaa
     7621 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7681 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7741 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     7801 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7861 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7921 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt
     7981 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     8041 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     8101 tgcgtcgaga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     8161 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     8221 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     8281 aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg
     8341 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     8401 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     8461 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     8521 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     8581 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     8641 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     8701 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     8761 ttacaatttc c
//
