LOCUS       sequence-237302-        3988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP L3-L2 entry plasmid, contains APEX2 + WPRE for 3- or 
            4-component MultiSite Gateway Pro assembly. Allows fusion of C-term 
            APEX2 to gene of interest in lentivirus vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3988)
  AUTHORS   Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, 
            Newgard CB
  TITLE     Creation of versatile cloning platforms for transgene expression and
            dCas9-based epigenome editing.
  JOURNAL   Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 
            10.1093/nar/gky1286.
  PUBMED    30590691
REFERENCE   2  (bases 1 to 3988)
  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: "Nucleic 
            Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286."
COMMENT     Sequence Label: JD602: pMVP (L3-L2) APEX2 + WPRE
FEATURES             Location/Qualifiers
     source          1..3988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             692..1435
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="KSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGTF
                     DKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAVE
                     VTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHTI
                     GAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDKY
                     AADEDAFFADYAEAHQKLSELGFADA"
     misc_feature    1484..2072
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1537..1557)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1955..1966)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2100..2199)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2229..2248)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2230..2248)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2253..2269)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2253..2269)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2371..2390)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2382..3191
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2449..2468)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3338..3926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3827..3846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa
      661 agttggtggc ggatccggcg gaggtggatc taagtcttac ccaactgtga gtgctgatta
      721 ccaggacgcc gttgagaagg cgaagaagaa gctcagaggc ttcatcgctg agaagagatg
      781 cgctcctcta atgctccgtt tggcattcca ctctgctgga acctttgaca agggcacgaa
      841 gaccggtgga cccttcggaa ccatcaagca ccctgccgaa ctggctcaca gcgctaacaa
      901 cggtcttgac atcgctgtta ggcttttgga gccactcaag gcggagttcc ctattttgag
      961 ctacgccgat ttctaccagt tggctggcgt tgttgccgtt gaggtcacgg gtggacctaa
     1021 ggttccattc caccctggaa gagaggacaa gcctgagccg ccaccagagg gtcgcttgcc
     1081 cgatcccact aagggttctg accatttgag agatgtgttt ggcaaagcta tggggcttac
     1141 tgaccaagat atcgttgctc tatctggggg tcacactatt ggagctgcac acaaggagcg
     1201 ttctggattt gagggtccct ggacctctaa tcctcttatt ttcgacaact catacttcac
     1261 ggagttgttg agtggtgaga aggaaggtct ccttcagcta ccttctgaca aggctctttt
     1321 gtctgaccct gtattccgcc ctctcgttga caaatatgca gcggacgaag atgccttctt
     1381 tgctgattac gctgaggctc accaaaagct ttccgagctt gggtttgctg atgcctaaga
     1441 attcctagag ctcctatagt gagaccggtt agtaatgatc gacaatcaac ctctggatta
     1501 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1561 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     1621 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     1681 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     1741 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     1801 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1861 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     1921 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     1981 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2041 gagtcggatc tccctttggg ccgcctcccc gcctggcgat ggtactgtcc tagtacctga
     2101 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2161 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg cagggcggcc
     2221 gcgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     2281 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     2341 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     2401 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     2461 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     2521 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2581 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2641 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     2701 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     2761 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     2821 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     2881 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     2941 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     3001 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     3061 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     3121 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     3181 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     3241 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     3301 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3421 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3481 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3541 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3601 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3661 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3721 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3781 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3841 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3901 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3961 ccttttgctg gccttttgct cacatgtt
//
