LOCUS       sequence-237287-        4341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP L3-L2 entry plasmid, contains Hygro-P2A + WPRE for 3- or 
            4-component MultiSite Gateway Pro assembly. Allows expression of 
            C-term Hygro selection marker linked by P2A to gene in lentivirus 
            vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4341)
  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 4341)
  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: LA301: pMVP (L3-L2) P2A-Hygro + WPRE
FEATURES             Location/Qualifiers
     source          1..4341
                     /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             680..736
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             743..1780
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1759..1767
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    1837..2425
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1890..1910)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2308..2319)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2453..2552)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2582..2601)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2583..2601)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2606..2622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2606..2622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2724..2743)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2735..3544
                     /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(2802..2821)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3691..4279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4180..4199
                     /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 ggatccggag ctactaattt ctctctgtta aagcaggcag gagatgtgga
      721 ggagaatcca ggtcctaagg gaatgaagaa gcccgaactc accgctacca gcgttgaaaa
      781 atttctcatc gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga
      841 gagccgagcc ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg
      901 cgctgatggc ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc
      961 catccccgaa gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag
     1021 acgcgcccaa ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca
     1081 gcctgtcgcc gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt
     1141 cggcccattc gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc
     1201 cattgctgat ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag
     1261 cgtagctcaa gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca
     1321 cctcgtccat gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc
     1381 cgtaatcgac tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt
     1441 cttctggcgg ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca
     1501 tcccgagctg gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca
     1561 gctctaccag agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg
     1621 cgatgccatc gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag
     1681 cgcagccgta tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc
     1741 cagtacacga ccgcgcgcta aggaggtagg tcgagtttaa gaattctgaa gcttagccta
     1801 gagctcctat agtgagaccg gttagtaatg atcgacaatc aacctctgga ttacaaaatt
     1861 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     1921 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     1981 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2041 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2101 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2161 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2221 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     2281 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2341 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     2401 atctcccttt gggccgcctc cccgcctggc gatggtactg tcctagtacc tgacccagct
     2461 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     2521 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagggcg gccgcgatat
     2581 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2641 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2701 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2761 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2821 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2881 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2941 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3001 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3061 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3121 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3181 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3241 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3301 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3361 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3421 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3481 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3541 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3601 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3661 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4081 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4261 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4321 ctggcctttt gctcacatgt t
//
