LOCUS       sequence-237170-        5718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMVP R4-R3 entry plasmid, contains beta-gal (no stop codon) 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 5718)
  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 5718)
  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: KH305: pMVP (R4-R3) beta-gal open
FEATURES             Location/Qualifiers
     source          1..5718
                     /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     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             735..3779
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVAPSNWQMHGYDAPIYSNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSETGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ASLQCTADTLADAALITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRPTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     protein_bind    3781..3904
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3959..3978)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3960..3978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3983..3999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3983..3999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4101..4120)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4112..4921
                     /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(4179..4198)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5068..5656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5557..5576
                     /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 aagttgtaac catggtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
      781 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
      841 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc
      901 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg
      961 tcgtcgcccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac tccaacgtga
     1021 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact
     1081 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg
     1141 atggcgttaa ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg
     1201 acagtcgttt gccgtctgaa tttgacctga gcgcattttt acgcgccgga gaaaaccgcc
     1261 tcgcggtgat ggtgctgcgc tggagtgacg gcagttatct ggaagatcag gatatgtggc
     1321 ggatgagcgg cattttccgt gacgtctcgt tgctgcataa accgactaca caaatcagcg
     1381 atttccatgt tgccactcgc tttaatgatg atttcagccg cgctgtactg gaggctgaag
     1441 ttcagatgtg cggcgagttg cgtgactacc tacgggtaac agtttcttta tggcagggtg
     1501 aaacgcaggt cgccagcggc accgcgcctt tcggcggtga aattatcgat gagcgtggtg
     1561 gttatgccga tcgcgtcaca ctacgtctga acgtcgaaaa cccgaaactg tggagcgccg
     1621 aaatcccgaa tctctatcgt gcggtggttg aactgcacac cgccgacggc acgctgattg
     1681 aagcagaagc ctgcgatgtc ggtttccgcg aggtgcggat tgaaaatggt ctgctgctgc
     1741 tgaacggcaa gccgttgctg attcgaggcg ttaaccgtca cgagcatcat cctctgcatg
     1801 gtcaggtcat ggatgagcag acgatggtgc aggatatcct gctgatgaag cagaacaact
     1861 ttaacgccgt gcgctgttcg cattatccga accatccgct gtggtacacg ctgtgcgacc
     1921 gctacggcct gtatgtggtg gatgaagcca atattgaaac ccacggcatg gtgccaatga
     1981 atcgtctgac cgatgatccg cgctggctac cggcgatgag cgaacgcgta acgcgaatgg
     2041 tgcagcgcga tcgtaatcac ccgagtgtga tcatctggtc gctggggaat gaatcaggcc
     2101 acggcgctaa tcacgacgcg ctgtatcgct ggatcaaatc tgtcgatcct tcccgcccgg
     2161 tgcagtatga aggcggcgga gccgacacca cggccaccga tattatttgc ccgatgtacg
     2221 cgcgcgtgga tgaagaccag cccttcccgg ctgtgccgaa atggtccatc aaaaaatggc
     2281 tttcgctacc tggagagacg cgcccgctga tcctttgcga atacgcccac gcgatgggta
     2341 acagtcttgg cggtttcgct aaatactggc aggcgtttcg tcagtatccc cgtttacagg
     2401 gcggcttcgt ctgggactgg gtggatcagt cgctgattaa atatgatgaa aacggcaacc
     2461 cgtggtcggc ttacggcggt gattttggcg atacgccgaa cgatcgccag ttctgtatga
     2521 acggtctggt ctttgccgac cgcacgccgc atccagcgct gacggaagca aaacaccagc
     2581 agcagttttt ccagttccgt ttatccgggc aaaccatcga agtgaccagc gaatacctgt
     2641 tccgtcatag cgataacgag ctcctgcact ggatggtggc gctggatggt aagccgctgg
     2701 caagcggtga agtgcctctg gatgtcgctc cacaaggtaa acagttgatt gaactgcctg
     2761 aactaccgca gccggagagc gccgggcaac tctggctcac agtacgcgta gtgcaaccga
     2821 acgcgaccgc atggtcagaa accgggcaca tcagcgcctg gcagcagtgg cgtctggcgg
     2881 aaaacctcag tgtgacgctc cccgccgcgt cccacgccat cccgcatctg accaccagcg
     2941 aaatggattt ttgcatcgag ctgggtaata agcgttggca atttaaccgc cagtcaggct
     3001 ttctttcaca gatgtggatt ggcgataaaa aacaactgct gacgccgctg cgcgatcagt
     3061 tcacccgtgc accgctggat aacgacattg gcgtaagtga agcgacccgc attgacccta
     3121 acgcctgggt cgaacgctgg aaggcggcgg gccattacca ggccgaagca gcgtcgttgc
     3181 agtgcacggc agatacactt gctgatgcgg cgctgattac gaccgctcac gcgtggcagc
     3241 atcaggggaa aaccttattt atcagccgga aaacctaccg gattgatggt agtggtcaaa
     3301 tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg cggattggcc
     3361 tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta gggccgcaag
     3421 aaaactatcc cgaccgccct actgccgcct gttttgaccg ctgggatctg ccattgtcag
     3481 acatgtatac cccgtacgtc ttcccgagcg aaaacggtct gcgctgcggg acgcgcgaat
     3541 tgaattatgg cccacaccag tggcgcggcg acttccagtt caacatcagc cgctacagtc
     3601 aacagcaact gatggaaacc agccatcgcc atctgctgca cgcggaagaa ggcacatggc
     3661 tgaatatcga cggtttccat atggggattg gtggcgacga ctcctggagc ccgtcagtat
     3721 cggcggaatt ccagctgagc gccggtcgct accattacca gttggtctgg tgtcaaaaga
     3781 caactttgta taataaagtt gaacgagaaa cgtaaaatga tataaatatc aatatattaa
     3841 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac
     3901 tatgaatcaa ctacttagat ggtattagtg acctgtactg cagggcggcc gcgatatccc
     3961 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     4021 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     4081 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     4141 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     4201 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     4261 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     4321 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     4381 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     4441 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     4501 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     4561 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     4621 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     4681 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     4741 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     4801 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     4861 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     4921 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     4981 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     5041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5461 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5521 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5581 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5641 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5701 gccttttgct cacatgtt
//
