LOCUS       sequence-356736-        7368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLenti vector backbone designed to express GFP from a 0.4CaMKIIa 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  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 7368)
  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: pJEP215-Lenti-0.4CaMKIIa P-GFP-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7368
                     /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    2326..2350
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2353..2362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2359..2361,2362..2364,2365..3078)
                     /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(2359..2361,2362..2364,2365..3075)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2404..2425)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2665..2684)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3012..3033
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3079..3103)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3156..3197
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3221..3809
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3274..3294)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3692..3703)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3889..4122
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4194..4315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4231..4250)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4285..4304
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4355..4490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4510..4529)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4511..4529)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4539..4556)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4539..4555)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4548..4570)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4697..5152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4784..4803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4994..5015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5178..5282
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5283..5351,5352..6143)
                     /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(5501..5520)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6314..6902
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6803..6822
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7056..7073
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7190..7211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7226..7231,7232..7249,7250..7256)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7264..7280
                     /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     7269..7291
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7288..7304
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7288..7304
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7323..7343
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7325..7343
                     /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 tatcgatact tgtggactaa gtttgttcac atccccttct ccaaccccct cagtacatca
     1981 ccctgggaga acaaggtcca cttgcttctg ggcccacaca gtcctgcagt attgtgtata
     2041 taaggccagg gcaacggagg agcaggtttt gaagtgaaag gcaggcaggt gttggggagg
     2101 cagttaccgg ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg
     2161 ctgacgaagg ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc
     2221 gtcaatcaag ctggttctcc atttgcactc aggagcacgg gcaggcgagt ggcccctagt
     2281 tctgggagca gactagtcca gtgtggtgga attctgcaga tatcaacaag tttgtacaaa
     2341 aaagcaggct acgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2401 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2461 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2521 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2581 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2641 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2701 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2761 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2821 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2881 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2941 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3001 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3061 gacgagctgt acaagtaaac ccagctttct tgtacaaagt ggttgatatc cagcacagtg
     3121 gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     3181 tcggtctcga ttctacgcgt accggttagt aatgatcgac aatcaacctc tggattacaa
     3241 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3301 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3361 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3421 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3481 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3541 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3601 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3661 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3721 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3781 tcggatctcc ctttgggccg cctccccgcc tggcgatggt acctttaaga ccaatgactt
     3841 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     3901 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     3961 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     4021 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     4081 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     4141 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     4201 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4261 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4321 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4381 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4441 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     4501 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     4561 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     4621 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     4681 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4741 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4801 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4861 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4921 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4981 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5041 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5101 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     5161 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5221 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5281 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5341 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5401 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5461 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5521 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5581 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5641 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5701 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5761 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5821 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5881 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5941 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6001 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6061 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6121 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6181 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6241 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6301 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6361 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6421 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6481 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6541 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6601 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6661 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6721 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6781 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6841 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6901 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6961 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7021 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7081 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     7141 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     7201 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     7261 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     7321 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagctt
//
