LOCUS       sequence-237083-        3370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP L5-L4 entry plasmid, contains BioID2 for 4-component MultiSite 
            Gateway Pro assembly. Allows fusion of N-term BioID2 to gene of 
            interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3370)
  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 3370)
  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: IO901: pMVP (L5-L4) BioID2
FEATURES             Location/Qualifiers
     source          1..3370
                     /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=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     5'UTR           680..722
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             734..1435
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="MAFKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRK
                     WLSQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKK
                     VSGVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRIS
                     ENLKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIK
                     EILSGEFSLRRS"
     protein_bind    complement(1485..1580)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1611..1630)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1612..1630)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1635..1651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1635..1651)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1753..1772)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1764..2573
                     /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(1831..1850)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2720..3308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3209..3228
                     /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 tgtatacaaa
      661 agttgtagaa tacaagcttc ttgttctttt tgcagaagct cagaataaac gctcaacttt
      721 ggcagatctg aacatggcct tcaagaacct gatctggctg aaggaggtgg acagcaccca
      781 ggagagactc aaggagtgga acgtgagcta cggcaccgcc cttgtggccg acagacagac
      841 caagggcaga ggcggcctgg gcagaaagtg gcttagccag gagggcggcc tgtacttcag
      901 cttcctgctc aaccccaagg agttcgagaa cctgcttcag ctgcccctcg tgcttggcct
      961 gagcgtgagc gaggccctgg aggagatcac cgagatcccc ttcagcctca agtggcccaa
     1021 cgacgtgtac ttccaggaga agaaggtgag cggcgtgctg tgcgagctta gcaaggacaa
     1081 gctgatcgtg ggcatcggca tcaacgtgaa ccagagagag atccccgagg agatcaagga
     1141 cagagccacc accctctacg agatcaccgg caaggactgg gacagaaagg aggtgctcct
     1201 taaggtgctg aagagaatca gcgagaacct caagaagttc aaggagaaga gcttcaagga
     1261 gttcaagggc aagatcgaga gcaagatgct gtacctgggc gaggaggtga agcttctggg
     1321 cgagggcaag atcaccggca agctggtggg ccttagcgag aagggcggcg ccctgatcct
     1381 gaccgaggag ggcatcaagg agatccttag cggcgagttc agcctgagaa gaagcggcgg
     1441 aggcggatct gggggtggag gctctggtgg agggggatcc cacccaactt ttctatacaa
     1501 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1561 tcaaaataaa atcattattt gccatccagc tgcagggcgg ccgcgatatc ccctatagtg
     1621 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     1681 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1741 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     1801 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1861 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1921 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1981 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     2041 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     2101 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     2161 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2221 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2281 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     2341 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2401 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2461 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2521 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     2581 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     2641 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     2701 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2761 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2821 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2881 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2941 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3001 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3061 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3121 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3181 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3241 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3301 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3361 ctcacatgtt
//
