LOCUS       sequence-356734-        8153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pLenti vector backbone containing a 1.3CaMKII promoter followed by 
            an intron followed by multiple cloning site(MCS) (reverse 
            orientation). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8153)
  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 8153)
  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: pJEP212-Lenti-1.3CaMKII-P-Intron-MCS-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..8153
                     /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"
     misc_feature    2055..2643
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2161..2172
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2570..2590
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     intron          2753..3228
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     promoter        complement(3308..4596)
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     LTR             4674..4907
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4979..5100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5070..5089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5140..5275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5295..5314)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5296..5314)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5324..5341)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5324..5340)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5333..5355)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5482..5937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5569..5588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5779..5800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5963..6067
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6068..6136,6137..6928)
                     /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(6286..6305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7099..7687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7588..7607
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7841..7858
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7975..7996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8011..8016,8017..8034,8035..8041)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8049..8065
                     /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     8054..8076
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8073..8089
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8073..8089
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8108..8128
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8110..8128
                     /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 tatcgatacg cgtcaattgg cggccgcctc gacagtgaaa aaaatgcttt atttgtgaaa
     1981 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaactc gagccacacc
     2041 ggtaccatcg ccaggcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag
     2101 gcgaagacgc ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg
     2161 attgagggcc gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacaca
     2221 ggcgagcagc catggaaagg acgtcagctt ccccgacaac accacggaat tgtcagtgcc
     2281 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc
     2341 aatagggagg gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca
     2401 accagtgggg gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg
     2461 ccacaactcc tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc
     2521 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
     2581 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
     2641 attgtcgaca tactagtatc ccgggatatg catgatgata tcatcatatg atgaattcac
     2701 gtgctttgcc aaagtgatgg gccagcacac agaccagcac gttgcccagg agctgtggga
     2761 ggaagataag aggtatgaac atgattagca aaagggccta gcttggactc agaataatcc
     2821 agccttatcc caaccataaa ataaaagcag aatggtagct ggattgtagc tgctattagc
     2881 aatatgaaac ctcttacatc agttacaatt tatatgcaga aatatttata tgcagaaata
     2941 ttgctattgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaggca
     3001 tgatacattg tatcattatt gccctgaaag aaagagatta gggaaagtat tagaaataag
     3061 ataaacaaaa aagtatatta aaagaagaaa gcatttttta aaattacaaa tgcaaaatta
     3121 ccctgatttg gtcaatatgt gtaccctgtt acttctcccc ttcctatgac atgaacttaa
     3181 ccatagaaaa gaaggggaaa gaaaacatca agggtcccat agactcaccc tgaagttctc
     3241 aggatccgag ctcggcgcgc cgctagcccg cggtcgcgaa cgcgtggccg gccgtagggg
     3301 atcccccgct gcccccagaa ctaggggcca ctcgcctgcc cgtgctcctg agtgcaaacg
     3361 gagaaccggc ttgactgacg agcttggggc ttctgagcag ggcactgtgg ctgctcacag
     3421 cctcgcgagc cttcgtcagc atccaggtcc ccatggcaac cacctccgaa acgccctgtt
     3481 cccgttgccc cggtaactgc ctccccaaca cctgcctgcc tttcacttta aaacctgctc
     3541 ctctttgccc tggccttata tacacaatac tgcaggactg tgtgggccca ggagcaagtg
     3601 gaccctgttc ccccagggtg atgtactgag ggggttggag aaggggatgc gaacaaactt
     3661 agtccacaag tgatcatcgc tcttcctacc tcccccacct ccaactatag ggccctttta
     3721 acagacggga tgggattaag tggggcaaag aggcagttgc tatggtaacg gctagtgggt
     3781 gccacattct gggatggtcc ttgagaaaga caggatgtag ttaccctggc aacttcatct
     3841 cctctgaaga gacaggtatt gccccgttgc cctggcaaca gctgggttct aaggtttaca
     3901 ggggaagcta attcatctct ccttcccttc ccccttgtag tctctgtaat ggtttctttg
     3961 cttcctctgg gaattggtct ggttgacttg gcctctgtct ctgtccttgt ttctttccag
     4021 ctctccctgc tttttagcca ttgtgcatat cacaccgcca ccttagcgtg agaagaagta
     4081 ccaaacagac ccagaatgct gacggaatga ctctgtctgc tttctcaccc ctatctgtca
     4141 ccactgagca gcagggcccc tcctgaggcc tctgccacag ctctccccac ccatcctccc
     4201 agagggacca tgaagatttg gtctcctggg gctcctgtcc cagtttctgc ttgtgcttcc
     4261 cgtcaaggta ggactggaga gaggcaacta taatgaggtg aacgtggctg gactcaccca
     4321 gggctgggcg ggtggtgtag atgtaccctg ccctggactc ttctttccta aactcattgg
     4381 tcatctgccc taatgagttc actccttctg tatatcgtac ttgcatgccc ccaaggcatg
     4441 gaatttaaga ttagtaatct gagttcaagt tctagctctg acatttctga tgtgaccctg
     4501 ggcaggtcac tgaggaagct ctctgcaccc ccatctcatt ttctagaaaa tcagaagaag
     4561 aaccccatgg agatgaagtg acctaaggcc ataatgttac gaggtacctt taagaccaat
     4621 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     4681 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     4741 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     4801 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     4861 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     4921 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     4981 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5041 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5101 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     5161 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5221 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     5281 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     5341 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5401 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5461 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5521 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5581 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5641 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5701 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5761 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5821 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5881 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     5941 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6001 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6061 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6121 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6181 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6241 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6301 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6361 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6421 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6481 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6541 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6601 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6661 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6721 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6781 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6841 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6901 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6961 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7021 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7081 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7141 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7201 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7261 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7321 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7381 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7441 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7501 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7561 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7621 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7681 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7741 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     7801 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7861 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     7921 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     7981 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     8041 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     8101 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag ctt
//
