LOCUS       sequence-356732-        7610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLenti vector backbone designed to express GFP from a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7610)
  AUTHORS   Holehonnur R, Lella SK, Ho A, Luong JA, Ploski JE
  TITLE     The production of viral vectors designed to express large and 
            difficult to express transgenes within neurons.
  JOURNAL   Mol Brain. 2015 Feb 24;8:12. doi: 10.1186/s13041-015-0100-7.
  PUBMED    25887710
REFERENCE   2  (bases 1 to 7610)
  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: "Mol 
            Brain."; date: "2015-02-24"; volume: "8:12. doi"; pages: " 
            10.1186/s13041-015-0100-7"
COMMENT     Sequence Label: pJEP217-Lenti-CMV-GFP-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1944..2247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2248..2451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2402..2422
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2448..2472
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2568..2592
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2595..2604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2601..2603,2604..2606,2607..3320)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2601..2603,2604..2606,2607..3317)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2646..2667)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2907..2926)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3254..3275
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3321..3345)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3398..3439
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3463..4051
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3516..3536)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3934..3945)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4131..4364
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4436..4557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4473..4492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4527..4546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4597..4732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4752..4771)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4753..4771)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4781..4798)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4781..4797)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4790..4812)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4939..5394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5026..5045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5236..5257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5420..5524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5525..5593,5594..6385)
                     /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(5743..5762)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6556..7144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7045..7064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7298..7315
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7432..7453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7468..7473,7474..7491,7492..7498)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7506..7522
                     /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     7511..7533
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7530..7546
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7530..7546
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7565..7585
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7567..7585
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1981 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2041 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2101 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2161 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2221 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2281 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2341 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2401 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     2461 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga
     2521 ctctagagga tccactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca
     2581 aaaaagcagg ctacgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2641 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2701 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2761 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2821 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2881 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2941 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3001 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3061 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3121 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3181 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3241 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3301 tggacgagct gtacaagtaa acccagcttt cttgtacaaa gtggttgata tccagcacag
     3361 tggcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     3421 cctcggtctc gattctacgc gtaccggtta gtaatgatcg acaatcaacc tctggattac
     3481 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3541 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3601 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3661 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3721 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3781 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3841 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     3901 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3961 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4021 agtcggatct ccctttgggc cgcctccccg cctggcgatg gtacctttaa gaccaatgac
     4081 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     4141 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     4201 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     4261 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     4321 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     4381 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     4441 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4501 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4561 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4621 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4681 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     4741 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     4801 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     4861 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4921 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4981 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5041 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5101 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5161 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5221 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5281 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5341 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     5401 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5521 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5581 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5641 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5701 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5761 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5821 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5881 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5941 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6001 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6061 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6121 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6181 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6241 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6301 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6361 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6421 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6481 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7201 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7261 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7321 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     7381 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     7441 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     7501 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     7561 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
//
