LOCUS       sequence-256179-        7530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of a peroxisomal eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7530)
  AUTHORS   Demaret T, Courtoy GE, Ravau J, Van Der Smissen P, Najimi M, Sokal 
            EM
  TITLE     Accurate and live peroxisome biogenesis evaluation achieved by 
            lentiviral expression of a green fluorescent protein fused to a 
            peroxisome targeting signal 1.
  JOURNAL   Histochem Cell Biol. 2020 Mar 2. pii: 10.1007/s00418-020-01855-z. 
            doi: 10.1007/s00418-020-01855-z.
  PUBMED    32124009
REFERENCE   2  (bases 1 to 7530)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Histochem 
            Cell Biol. 2020 Mar 2. pii: 10.1007/s00418-020-01855-z. doi: 
            10.1007/s00418-020-01855-z."
COMMENT     Sequence Label: pRRLSIN.cPPT.PGK-GFP-PTS1.WPRE
FEATURES             Location/Qualifiers
     source          1..7530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /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"
     misc_feature    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1972..2482
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2401..2419
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      2498..2507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2504..2506,2507..2509,2510..3220)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2504..2506,2507..2509,2510..3220)
                     /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(2549..2570)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2810..2829)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3157..3178
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3381..3969
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3434..3454)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3852..3863)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4056..4289
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4361..4482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4398..4417)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4452..4471
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4522..4657
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4677..4696)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4678..4696)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4706..4723)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4706..4722)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4715..4737)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4864..5319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4951..4970)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5161..5182
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5345..5449
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5450..5518,5519..6310)
                     /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     complement(5668..5687)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6481..7069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6970..6989
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7223..7240
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7357..7378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7393..7398,7399..7416,7417..7423)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7431..7447
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     7436..7458
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7455..7471
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7455..7471
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7490..7510
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7492..7510
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttatcg atcacgagac tagcctcgag aagcttgata tcgaattcca cggggttggg
     1981 gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg
     2041 ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag
     2101 cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc
     2161 cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc
     2221 acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc
     2281 gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga
     2341 aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg
     2401 gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg
     2461 aatcaccgac ctctctcccc agggggatcc accggtcgcc accatggtga gcaagggcga
     2521 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2581 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2641 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2701 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2761 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2821 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2881 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2941 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3001 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3061 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3121 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3181 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc
     3241 tcaagcttcg aattctaggt ccaagttata aggatccacc ggatctagat aactgatcat
     3301 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3361 cctgaacctg aaacgtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3421 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3481 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3541 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3601 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3661 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3721 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3781 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3841 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3901 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3961 cctccccgcc tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt
     4021 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     4081 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     4141 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     4201 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     4261 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     4321 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     4381 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4441 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     4501 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4561 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4621 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     4681 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     4741 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     4801 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4861 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4921 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4981 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5041 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5101 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5161 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5221 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5281 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     5341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5461 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5521 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5581 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5641 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5701 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5761 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5821 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5881 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5941 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6001 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6061 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6121 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6181 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6241 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6301 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6361 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6421 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6481 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6541 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6601 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6661 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6721 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6781 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6841 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6901 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6961 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7021 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7081 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7141 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     7201 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     7261 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     7321 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     7381 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     7441 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     7501 tcactaaagg gaacaaaagc tggagctgca
//
