LOCUS       sequence-237301-        3955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP L3-L2 entry plasmid, contains BioID2 + WPRE for 3- or 
            4-component MultiSite Gateway Pro assembly. Allows fusion of C-term 
            BioID2 to gene of interest in lentivirus vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3955)
  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 3955)
  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: JD404: pMVP (L3-L2) BioID2 + WPRE
FEATURES             Location/Qualifiers
     source          1..3955
                     /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             707..1402
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     misc_feature    1451..2039
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1504..1524)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1922..1933)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2067..2166)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2196..2215)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2197..2215)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2220..2236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2220..2236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2338..2357)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2349..3158
                     /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(2416..2435)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3305..3893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3794..3813
                     /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 tggaggtgga ggctccttca agaacctgat
      721 ctggctgaag gaggtggaca gcacccagga gagactcaag gagtggaacg tgagctacgg
      781 caccgccctt gtggccgaca gacagaccaa gggcagaggc ggcctgggca gaaagtggct
      841 tagccaggag ggcggcctgt acttcagctt cctgctcaac cccaaggagt tcgagaacct
      901 gcttcagctg cccctcgtgc ttggcctgag cgtgagcgag gccctggagg agatcaccga
      961 gatccccttc agcctcaagt ggcccaacga cgtgtacttc caggagaaga aggtgagcgg
     1021 cgtgctgtgc gagcttagca aggacaagct gatcgtgggc atcggcatca acgtgaacca
     1081 gagagagatc cccgaggaga tcaaggacag agccaccacc ctctacgaga tcaccggcaa
     1141 ggactgggac agaaaggagg tgctccttaa ggtgctgaag agaatcagcg agaacctcaa
     1201 gaagttcaag gagaagagct tcaaggagtt caagggcaag atcgagagca agatgctgta
     1261 cctgggcgag gaggtgaagc ttctgggcga gggcaagatc accggcaagc tggtgggcct
     1321 tagcgagaag ggcggcgccc tgatcctgac cgaggagggc atcaaggaga tccttagcgg
     1381 cgagttcagc ctgagaagaa gctaagaatt cctagagctc ctatagtgag accggttagt
     1441 aatgatcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     1501 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     1561 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     1621 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     1681 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     1741 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     1801 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     1861 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     1921 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     1981 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     2041 tggcgatggt actgtcctag tacctgaccc agctttcttg tacaaagttg gcattataag
     2101 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2161 tatttgccat ccagctgcag ggcggccgcg atatccccta tagtgagtcg tattacatgg
     2221 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     2281 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     2341 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     2401 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     2461 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     2521 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     2581 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     2641 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     2701 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     2761 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     2821 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     2881 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     2941 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     3001 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     3061 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     3121 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     3181 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     3241 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3301 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3361 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3421 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3481 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3541 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3601 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3661 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3721 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3781 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3841 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3901 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
