LOCUS       sequence-237300-        3573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP L3-L2 entry plasmid, contains APEX2 + polyA for 3- or 
            4-component MultiSite Gateway Pro assembly. Allows fusion of C-term 
            APEX2 to gene of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3573)
  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 3573)
  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: IY202: pMVP (L3-L2) APEX2 + polyA
FEATURES             Location/Qualifiers
     source          1..3573
                     /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"
     primer_bind     complement(1463..1480)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1469..1676
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(1685..1784)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1814..1833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1815..1833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1838..1854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1838..1854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1956..1975)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1967..2776
                     /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(2034..2053)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2923..3511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3412..3431
                     /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 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1501 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1561 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1621 gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc tggggaggta
     1681 cctgacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     1741 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcaggg
     1801 cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1861 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1921 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1981 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2041 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2101 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2161 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2221 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2281 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2341 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2401 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2461 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2521 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2581 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2641 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2701 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2761 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2821 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3541 cctggccttt tgctggcctt ttgctcacat gtt
//
