LOCUS       sequence-255819-        4443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP R4-R3 entry plasmid, contains destabilized Firefly Luciferase 
            (Luc2P) for 3- or 4-component MultiSite Gateway Pro assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  TITLE     Unpublished pMVP and pMAGIC plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4443)
  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: 
            "Unpublished"
COMMENT     Sequence Label: LO601: pMVP (R4-R3) destabilized firefly lucif...
FEATURES             Location/Qualifiers
     source          1..4443
                     /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    complement(603..726)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             729..2378
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2382..2501
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     protein_bind    2506..2629
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2684..2703)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2685..2703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2708..2724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2708..2724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2826..2845)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2837..3646
                     /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(2904..2923)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3793..4381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4282..4301
                     /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 tcgggcccct acaggtcact aataccatct aagtagttga
      601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
      661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca
      721 aagttgccat ggaagatgcc aaaaacatta agaagggccc agcgccattc tacccactcg
      781 aagacgggac cgccggcgag cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg
      841 gcaccatcgc ctttaccgac gcacatatcg aggtggacat tacctacgcc gagtacttcg
      901 agatgagcgt tcggctggca gaagctatga agcgctatgg gctgaataca aaccatcgga
      961 tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc cgtgttgggt gccctgttca
     1021 tcggtgtggc tgtggcccca gctaacgaca tctacaacga gcgcgagctg ctgaacagca
     1081 tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa agggctgcaa aagatcctca
     1141 acgtgcaaaa gaagctaccg atcatacaaa agatcatcat catggatagc aagaccgact
     1201 accagggctt ccaaagcatg tacaccttcg tgacttccca tttgccaccc ggcttcaacg
     1261 agtacgactt cgtgcccgag agcttcgacc gggacaaaac catcgccctg atcatgaaca
     1321 gtagtggcag taccggattg cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc
     1381 gattcagtca tgcccgcgac cccatcttcg gcaaccagat catccccgac accgctatcc
     1441 tcagcgtggt gccatttcac cacggcttcg gcatgttcac cacgctgggc tacttgatct
     1501 gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga gctattcttg cgcagcttgc
     1561 aagactataa gattcaatct gccctgctgg tgcccacact atttagcttc ttcgctaaga
     1621 gcactctcat cgacaagtac gacctaagca acttgcacga gatcgccagc ggcggggcgc
     1681 cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc
     1741 agggctacgg cctgacagaa acaaccagcg ccattctgat cacccccgaa ggggacgaca
     1801 agcctggcgc agtaggcaag gtggtgccct tcttcgaggc taaggtggtg gacttggaca
     1861 ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca
     1921 tgagcggcta cgttaacaac cccgaggcta caaacgctct catcgacaag gacggctggc
     1981 tgcacagcgg cgacatcgcc tactgggacg aggacgagca cttcttcatc gtggaccggc
     2041 tgaagagcct gatcaaatac aagggctacc aggtagcccc agccgaactg gagagcatcc
     2101 tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg
     2161 gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga
     2221 tcgtggacta tgtggccagc caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt
     2281 tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga cgcccgcaag atccgcgaga
     2341 ttctcattaa ggccaagaag ggcggcaaga tcgccgtgaa ttctcacggc ttccctcccg
     2401 aggtggagga gcaggccgcc ggcaccctgc ccatgagctg cgcccaggag agcggcatgg
     2461 atagacaccc tgctgcttgc gccagcgcca ggatcaacgt ctaagcaact ttgtataata
     2521 aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     2581 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatga atcaactact
     2641 tagatggtat tagtgacctg tactgcaggg cggccgcgat atcccctata gtgagtcgta
     2701 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2761 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2821 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2881 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2941 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3001 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3061 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3121 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3181 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3241 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3301 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3361 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3421 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3481 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3541 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3601 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3661 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3721 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4081 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4141 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4201 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4261 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4321 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4381 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4441 gtt
//
