LOCUS       sequence-171187-        9089 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector for CMV promoter driven expression of EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9089)
  AUTHORS   Ou W, Marino MP, Suzuki A, Joshi B, Husain SR, Maisner A, Galanis E,
            Puri RK, Reiser J
  TITLE     Specific targeting of human interleukin (IL)-13 receptor 
            alpha2-positive cells with lentiviral vectors displaying IL-13.
  JOURNAL   Hum Gene Ther Methods. 2012 Apr;23(2):137-47. Epub 2012 May 21.
  PUBMED    22612657
REFERENCE   2  (bases 1 to 9089)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Hum Gene 
            Ther Methods."; date: "2012-04"; volume: "23(2)"; pages: "137-47. 
            Epub 2012 May 21"
COMMENT     Sequence Label: pNL(CMV)EGFP/CMV/WPRE∆U3
FEATURES             Location/Qualifiers
     source          1..9089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             586..766
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      1393..1528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    1806..2039
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2224..2268
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     enhancer        2538..2841
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2842..3045
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3084..3093
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3090..3092,3093..3095,3096..3806)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3090..3092,3093..3095,3096..3806)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3135..3156)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3396..3415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3743..3764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3826..4414
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3879..3899)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4297..4308)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4489..4722
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6362..6379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6533..7121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6613..6632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7292..8083,8084..8152))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7915..7934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8153..8257)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8325..8343
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8381..8403)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8503..8522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tggcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatggaatg gatgaccctg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtatcta gatggagttc cgcgttacat aacttacggt aaatggcccg
      361 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      421 gtaacgccaa tagggacttt ccattgacgt caatgggagt ttgttttggc accaaaatca
      481 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
      541 tgtactctag atgctacata taagcagctg ctttttgcct gtactgggtc tctctggtta
      601 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      661 taaagcttgc cttgagtgct caaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      721 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
      781 gggacttgaa agcgaaagta aagccagagg agatctctcg acgcaggact cggcttgctg
      841 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
      901 gcggaggcta gaaggagaga gatgggtgcg agagcgtcgg tattaagcgg gggagaatta
      961 gataaatggg aaaaaattcg gttaaggcca gggggaaaga aacaatataa actaaaacat
     1021 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct tttagagaca
     1081 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1141 gaacttagat cattatataa tacaatagca gtcctctatt gtgtgcatca aaggatagat
     1201 gtaaaagaca ccaaggaagc cttagataag atagaggaag agcaaaacaa aagtaagaaa
     1261 aaggcacagc aagcagcagc tgacacagga aacaacagcc aggtcagcca aaattaccct
     1321 atagtgcaga acctccaggg gcaaatggta catcaggcca tatcacctag aactttaaat
     1381 atgactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     1441 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     1501 aagtagtgag gaggcttttt tggaggccta ggcttttgca aatgcataaa agaaaagggg
     1561 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     1621 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     1681 gcagagatcc acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1741 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1801 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1861 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1921 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1981 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     2041 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     2101 tggagtaata aatctctgga acagatttgg aataacatga cctggatgga gtgggacaga
     2161 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2221 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2281 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2341 ggtttaagaa tagtttttgc tgtactttct gtagtgaata gagttaggca gggatattca
     2401 ccattatcgt ttcagaccca cctcccaatc ccgaggggac ccgacaggcc cgaaggaata
     2461 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctagt
     2521 taactagatc tagagtccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     2581 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     2641 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     2701 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     2761 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     2821 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     2881 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     2941 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     3001 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     3061 cagatccgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     3121 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3181 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3241 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3301 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3361 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3421 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3481 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3541 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3601 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3661 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3721 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3781 ctctcggcat ggacgagctg tacaagtccg gactcagatc tcgagaatca acctctggat
     3841 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3901 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3961 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     4021 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     4081 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     4141 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4201 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     4261 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4321 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4381 acgagtcgga tctccctttg ggccgcctcc ccgcctgggt acctttaaga ccaatgactt
     4441 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     4501 ttcactccca aagaagacaa gatctgcttt ttgcctgtac tgggtctctc tggttagacc
     4561 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     4621 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     4681 gatccctcag acccttttag tcagtgtgga aaatctctag cacccaggag gtagaggttg
     4741 cagtgagcca agatcgcgcc actgcattcc agcctgggca agaaaacaag actgtctaaa
     4801 ataataataa taagttaagg gtattaaata tatttataca tggaggtcat aaaaatatat
     4861 atatttgggc tgggcgcagt ggctcacacc tgcgcccggc cctttgggag gccgaggcag
     4921 gtggatcacc tgagtttggg agttccagac cagcctgacc aacatggaga aaccccttct
     4981 ctgtgtattt ttagtagatt ttattttatg tgtattttat tcacaggtat ttctggaaaa
     5041 ctgaaactgt ttttcctcta ctctgatacc acaagaatca tcagcacaga ggaagacttc
     5101 tgtgatcaaa tgtggtggga gagggaggtt ttcaccagca catgagcagt cagttctgcc
     5161 gcagactcgg cgggtgtcct tcggttcagt tccaacaccg cctgcctgga gagaggtcag
     5221 accacagggt gagggctcag tccccaagac ataaacaccc aagacataaa cacccaacag
     5281 gtccaccccg cctgctgccc aggcagagcc gattcaccaa gacgggaatt aggatagaga
     5341 aagagtaagt cacacagagc cggctgtgcg ggagaacgga gttctattat gactcaaatc
     5401 agtctcccca agcattcggg gatcagagtt tttaaggata acttagtgtg tagggggcca
     5461 gtgagttgga gatgaaagcg tagggagtcg aaggtgtcct tttgcgccga gtcagttcct
     5521 gggtgggggc cacaagatcg gatgagccag tttatcaatc cgggggtgcc agctgatcca
     5581 tggagtgcag ggtctgcaaa atatctcaag cactgattga tcttaggttt tacaatagtg
     5641 atgttacccc aggaacaatt tggggaaggt cagaatcttg tagcctgtag ctgcatgact
     5701 cctaaaccat aatttctttt ttgttttttt ttttttattt ttgagacagg gtctcactct
     5761 gtcacctagg ctggagtgca gtggtgcaat cacagctcac tgcagcctca acgtcgtaag
     5821 ctcaagcgat cctcccacct cagcctgcct ggtagctgag actacaagcg acgccccagt
     5881 taatttttgt atttttggta gaggcagcgt tttgccgtgt ggccctggct ggtctcgaac
     5941 tcctgggctc aagtgatcca gcctcagcct cccaaagtgc tgggacaacc ggggccagtc
     6001 actgcacctg gccctaaacc ataatttcta atcttttggc taatttgtta gtcctacaaa
     6061 ggcagtctag tccccaggca aaaagggggt ttgtttcggg aaagggctgt tactgtcttt
     6121 gtttcaaact ataaactaag ttcctcctaa acttagttcg gcctacaccc aggaatgaac
     6181 aaggagagct tggaggttag aagcacgatg gaattggtta ggtcagatct ctttcactgt
     6241 ctgagttata attttgcaat ggtggttcaa agactgcccg cttctgacac cagtcgctgc
     6301 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6361 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6421 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6481 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6541 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6601 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6661 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6721 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6781 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6841 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6901 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6961 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7021 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7081 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7141 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7201 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7261 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7321 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7381 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7441 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7501 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7561 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7621 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7681 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7741 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7801 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7861 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7921 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7981 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8041 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8101 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8161 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8221 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8281 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     8341 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     8401 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     8461 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     8521 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     8581 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     8641 cctcttcgct attacgccag gggaggcaga gattgcagta agctgagatc gcagcactgc
     8701 actccagcct gggcgacaga gtaagactct gtctcaaaaa taaaataaat aaatcaatca
     8761 gatattccaa tcttttcctt tatttattta tttattttct attttggaaa cacagtcctt
     8821 ccttattcca gaattacaca tatattctat ttttctttat atgctccagt tttttttaga
     8881 ccttcacctg aaatgtgtgt atcgagacct agaaaaacat ggagcaatca caagtagcaa
     8941 tacagcagct aacaatgctg cttgtgcctg gctagaagca caagaggagg aagaggtggg
     9001 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     9061 tagccacttt ttaaaagaaa aggggggac
//
