LOCUS       sequence-237514-        4520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP L3-L2 entry plasmid, contains HA tag-polyA + RIP-TETa-polyA for
            3- or 4-component MultiSite Gateway Pro assembly. Adds C-term HA tag
            to gene plus downstream RIP-driven TETa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4520)
  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 4520)
  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: KO502: pMVP (L3-L2) HA tag + pA; RIP::TETa-pA
FEATURES             Location/Qualifiers
     source          1..4520
                     /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             686..712
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    845..926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             join(1390..2007,2008..2013,2014..2049,2050..2052,2053..2088,
                     2089..2091,2092..2127,2128..2136)
                     /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    2159..2280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2170..2189)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2632..2731)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2761..2780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2762..2780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2785..2801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2785..2801)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2903..2922)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2914..3723
                     /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(2981..3000)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3870..4458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4359..4378
                     /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 aacaaagctg gatcctaccc atacgatgtg ccagattacg cttgatttaa
      721 atagataact gactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
      781 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
      841 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      901 cacaaataaa gcattttttt cactgcctcg agtctagact gagcaagaat ccagctatca
      961 atagaaacta tgaaacagtt ccagggacaa agataccagg tccccaacaa ctgcaacttt
     1021 ctgggaaatg aggtggaaaa tgctcagcca aggaaaaaga gggccttacc ctctctggga
     1081 caatgattgt gctgtgaact gcttcatcag gccatctggc cccttgttaa taatctaatt
     1141 accctaggtc taagtagagt tgttgacgtc caatgagcgc tttctgcaga cttagcacta
     1201 ggcaagtgtt tggaaattac agcttcagcc cctctcgcca tctgcctacc tacccctcct
     1261 agagccctta atgggccaaa cggcaaagtc cagggggcag agaggaggtg ctttggacta
     1321 taaagctagt ggagacccag taactcccaa ccctaagtga ccagctacga attcgactgg
     1381 cgcgccacca tgtccagact ggacaagagc aaagtcataa acggcgctct ggaattactc
     1441 aatggagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     1501 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     1561 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     1621 tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct
     1681 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     1741 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     1801 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     1861 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     1921 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     1981 gagaaacagc taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta
     2041 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     2101 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaagaatcc agacatgata
     2161 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2221 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2281 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2341 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctgc aagcctcgtc
     2401 gtcctggccg gaccacgcta tctgtgcaag gtccccggcc ccggacgcgc gctccatgag
     2461 cagagcgccc gccgccgagg cgaagactcg ggcggcgccc tgcccgtccc accaggtcaa
     2521 caggcggtaa ccggcctctt catcgggaat gcgcgcgacc ttcagcatcg ccggcatgtc
     2581 cccctggcgg acgggaagta tccagctcga ccaggcttgt cgactcgaag aacccagctt
     2641 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     2701 tcactatcag tcaaaataaa atcattattt gccatccagc tgcagggcgg ccgcgatatc
     2761 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2821 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     2881 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2941 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     3001 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     3061 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     3121 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     3181 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     3241 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     3301 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     3361 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     3421 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     3481 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     3541 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     3601 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3661 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3721 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3781 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3841 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4441 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4501 tggccttttg ctcacatgtt
//
