LOCUS       sequence-237309-        4671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP L3-L2 entry plasmid, contains polyA + CMV-TETa-polyA for 3- or 
            4-component MultiSite Gateway Pro assembly. Adds CMV-driven TETa 
            downstream of gene of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4671)
  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 4671)
  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: KA501: pMVP (L3-L2) pA; CMV::TETa-pA
FEATURES             Location/Qualifiers
     source          1..4671
                     /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"
     polyA_signal    820..901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        965..1268
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1269..1472
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1423..1443
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1541..2158,2159..2164,2165..2200,2201..2203,2204..2239,
                     2240..2242,2243..2278,2279..2287)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    2310..2431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2321..2340)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2783..2882)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2912..2931)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2913..2931)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2936..2952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2936..2952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3054..3073)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3065..3874
                     /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(3132..3151)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4021..4609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4510..4529
                     /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 agttgagatc aacaaagctt cctagctcga tttaaataga taactgacta gaggatcata
      721 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
      781 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
      841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
      901 cctcgagtct agaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      961 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     1021 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     1081 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     1141 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1201 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     1261 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     1321 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     1381 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     1441 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
     1501 gctaccggac tcagatctcg aattcgactg gcgcgccacc atgtccagac tggacaagag
     1561 caaagtcata aacggcgctc tggaattact caatggagtc ggtatcgaag gcctgacgac
     1621 aaggaaactc gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa
     1681 caagcgggcc ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt
     1741 ctgccccctg gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcattccg
     1801 ctgtgctctc ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga
     1861 gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct
     1921 ggagaacgca ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga
     1981 ggaacaggag catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc
     2041 cccacttctg agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct
     2101 tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg
     2161 gccggccgac gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga
     2221 ctttgacctt gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc
     2281 cgggtaacta agtaagaatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     2341 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2401 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2461 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2521 tatggctgat tatgatcctg caagcctcgt cgtcctggcc ggaccacgct atctgtgcaa
     2581 ggtccccggc cccggacgcg cgctccatga gcagagcgcc cgccgccgag gcgaagactc
     2641 gggcggcgcc ctgcccgtcc caccaggtca acaggcggta accggcctct tcatcgggaa
     2701 tgcgcgcgac cttcagcatc gccggcatgt ccccctggcg gacgggaagt atccagctcg
     2761 accaggcttg tcgactcgaa gaacccagct ttcttgtaca aagttggcat tataagaaag
     2821 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     2881 tgccatccag ctgcagggcg gccgcgatat cccctatagt gagtcgtatt acatggtcat
     2941 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     3001 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     3061 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     3121 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     3181 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     3241 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     3301 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     3361 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3421 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3481 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3541 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3601 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3661 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3721 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3781 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3841 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3901 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3961 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4141 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
