LOCUS       sequence-356739-        7544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLenti vector backbone designed to express GFP from a 0.5Synapsin 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7544)
  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 7544)
  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: pJEP214-Lenti-0.5Synapsin P-GFP- WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7544
                     /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"
     protein_bind    2505..2529
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2532..2541
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2538..2540,2541..2543,2544..3257)
                     /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(2538..2540,2541..2543,2544..3254)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2583..2604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2844..2863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3191..3212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3258..3282)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3335..3376
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3400..3988
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3453..3473)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3871..3882)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4068..4301
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4373..4494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4410..4429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4464..4483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4534..4669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4686..4705)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4687..4705)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4715..4732)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4715..4731)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4724..4746)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4873..5328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4960..4979)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5170..5191
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5354..5458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5459..5527,5528..6319)
                     /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(5677..5696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6490..7078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6979..6998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7232..7249
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7366..7387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7402..7407,7408..7425,7426..7432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7440..7456
                     /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     7445..7467
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7464..7480
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7464..7480
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7499..7519
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7501..7519
                     /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 tatcgataca agtatctggg aagggtaaca atgcaagtta gcctgaggat ttaggaggag
     1981 gctgaaaaac agagtaggag ccttactacg ggtccagacc ctacggacaa gaacccccac
     2041 tcccactccc caaattgcgc attccctccc ccatcagagg gggaggggaa gaggatgcag
     2101 cgcggcgcgg cgcgtgcgca ctgtcggatt tagtaccgcg gacagagcct tcgcccccgc
     2161 tgccggcgcg cgccaccacc tccccagcac caaaggcggg ctgacgtcac tctccagccc
     2221 tccccaaact cccctacctc accgccttgg tcgcgtccgt gcagcggtga gtccagtcgg
     2281 gccgcaccac aagaggtgca agataggggg gtgcaggcgc gaccatacgc tctgcggcgg
     2341 cagagcctca gcgctgcctc agtctgcagc gggcagcaga ggagtcgcgt cgtgccagag
     2401 agcgccgccg tgctcctgag ccccttgcgc tccgcccccg cggcccaccg acccactgcc
     2461 ccttggatcc actagtccag tgtggtggaa ttctgcagat atcaacaagt ttgtacaaaa
     2521 aagcaggcta cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2581 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2641 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2701 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2761 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2821 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2881 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2941 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3001 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3061 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3121 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3181 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3241 acgagctgta caagtaaacc cagctttctt gtacaaagtg gttgatatcc agcacagtgg
     3301 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     3361 cggtctcgat tctacgcgta ccggttagta atgatcgaca atcaacctct ggattacaaa
     3421 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3481 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3541 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3601 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3661 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3721 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3781 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3841 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3901 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3961 cggatctccc tttgggccgc ctccccgcct ggcgatggta cctttaagac caatgactta
     4021 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     4081 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     4141 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     4201 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     4261 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     4321 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     4381 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4441 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4501 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4621 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac
     4681 ccaagcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     4741 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4801 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     4861 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4921 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4981 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5041 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5101 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5161 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5221 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5281 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     5341 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5401 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5461 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5521 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5581 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5641 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5701 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5761 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5821 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5881 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5941 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6001 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6061 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6121 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6181 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6241 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6301 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6361 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6421 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6481 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6541 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6601 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6661 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6721 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6781 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6841 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6901 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6961 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7021 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7081 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     7141 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7201 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7261 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     7321 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     7381 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     7441 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
     7501 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gctt
//
