LOCUS       sequence-336789-        6565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for CMV promoter driven expression of dominant 
            negative (DN) MAML1 (Notch pathway) fused to mVenus fluorescent 
            protein (to be used in conjunction with Phoenix packaging cells)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  AUTHORS   Hoare M, Ito Y, Kang TW, Weekes MP, Matheson NJ, Patten DA, Shetty 
            S, Parry AJ, Menon S, Salama R, Antrobus R, Tomimatsu K, Howat W, 
            Lehner PJ, Zender L, Narita M
  TITLE     NOTCH1 mediates a switch between two distinct secretomes during 
            senescence.
  JOURNAL   Nat Cell Biol. 2016 Sep;18(9):979-92. doi: 10.1038/ncb3397. Epub 
            2016 Aug 15.
  PUBMED    27525720
REFERENCE   2  (bases 1 to 6565)
  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"; doi: "10.1038/ncb3397"; 
            journalName: "Nat Cell Biol"; date: "2016-09"; volume: "18"; issue: 
            "9"; pages: "979-92"
COMMENT     Sequence Label: pLPC-dnMAML1-mVenus
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1369..1968
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1369..1388)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1865..1885
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2057..2076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2248..2627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2628..2831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2782..2802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2876..2895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2876..2894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2926..2935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3145..3147,3148..3150,3151..3864)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3190..3211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3451..3470)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3798..3819
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4052..4521
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4651..4668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4822..5410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4902..4921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5570..6361,6362..6430))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6193..6212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6431..6535)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc agcttaccat gaccgagtac
     1381 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     1441 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     1501 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     1561 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     1621 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     1681 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     1741 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     1801 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     1861 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     1921 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     1981 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggatct cccgatcccc
     2041 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     2101 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     2161 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     2221 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     2281 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2341 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2401 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2461 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2521 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2581 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     2641 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     2701 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2761 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2821 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
     2881 cgactcacta tagggagacc caagcttggt accgagctcg gatccgccac catggcgctg
     2941 ccgcggcaca gcgcggtcat ggagcgcctt cgccggcgca tcgagctgtg ccggcgccac
     3001 cacagcacct gcgaggcccg ctacgaggcc gtgtcgcccg agcgcctgga gctggagcgc
     3061 caacacacct tcgccctgca ccagcgctgc atccaggcca aggccaagcg cgccgggaag
     3121 cacgggaatt ctggcagatc tatcatggtg agcaagggcg aggagctgtt caccggggtg
     3181 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3241 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     3301 aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc
     3361 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3421 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3481 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3541 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3601 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc
     3661 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3721 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc
     3781 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3841 ggcatggacg agctgtacaa gtaagtcgag catgcatcta gagggccgat aaaataaaag
     3901 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     3961 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     4021 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     4081 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     4141 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     4201 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     4261 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     4321 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     4381 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     4441 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     4501 cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt
     4561 aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg
     4621 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4681 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4741 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4801 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4861 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4921 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4981 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5041 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5101 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5161 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5221 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5281 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5341 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5401 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5521 gcggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5581 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5641 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5701 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5761 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5821 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5881 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5941 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6001 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6061 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6121 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6181 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6241 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6301 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6361 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6421 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6541 ttccccgaaa agtgccacct gcagc
//
