LOCUS       sequence-338649-        8053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for expression of NOTCH1 intracellular domain with
            a C-terminal FLAG tag (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 8053)
  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 8053)
  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: pWZL-N1ICD-Flag
FEATURES             Location/Qualifiers
     source          1..8053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3772..3795
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3867..4328
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3923..3940)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4150..4169
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4333..5358
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4333..5358
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             5547..6016
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6146..6163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6317..6905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6397..6416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7065..7856,7857..7925))
                     /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     7688..7707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7926..8030)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccgc caccatgcgc
     1381 aagcgccggc ggcagcatgg ccagctctgg ttccctgagg gcttcaaagt gtctgaggcc
     1441 agcaagaaga agcggcggga gcccctcggc gaggactccg tgggcctcaa gcccctgaag
     1501 aacgcttcag acggtgccct catggacgac aaccagaatg agtgggggga cgaggacctg
     1561 gagaccaaga agttccggtt cgaggagccc gtggttctgc ctgacctgga cgaccagaca
     1621 gaccaccggc agtggactca gcagcacctg gatgccgctg acctgcgcat gtctgccatg
     1681 gcccccacac cgccccaggg tgaggttgac gccgactgca tggacgtcaa tgtccgcggg
     1741 cctgatggct tcaccccgct catgatcgcc tcctgcagcg ggggcggcct ggagacgggc
     1801 aacagcgagg aagaggagga cgcgccggcc gtcatctccg acttcatcta ccagggcgcc
     1861 agcctgcaca accagacaga ccgcacgggc gagaccgcct tgcacctggc cgcccgctac
     1921 tcacgctctg atgccgccaa gcgcctgctg gaggccagcg cagatgccaa catccaggac
     1981 aacatgggcc gcaccccgct gcatgcggct gtgtctgccg acgcacaagg tgtcttccag
     2041 atcctgatcc ggaaccgagc cacagacctg gatgcccgca tgcatgatgg cacgacgcca
     2101 ctgatcctgg ctgcccgcct ggccgtggag ggcatgctgg aggacctcat caactcacac
     2161 gccgacgtca acgccgtaga tgacctgggc aagtccgccc tgcactgggc cgccgccgtg
     2221 aacaatgtgg atgccgcagt tgtgctcctg aagaacgggg ctaacaaaga tatgcagaac
     2281 aacagggagg agacacccct gtttctggcc gcccgggagg gcagctacga gaccgccaag
     2341 gtgctgctgg accactttgc caaccgggac atcacggatc atatggaccg cctgccgcgc
     2401 gacatcgcac aggagcgcat gcatcacgac atcgtgaggc tgctggacga gtacaacctg
     2461 gtgcgcagcc cgcagctgca cggagccccg ctggggggca cgcccaccct gtcgcccccg
     2521 ctctgctcgc ccaacggcta cctgggcagc ctcaagcccg gcgtgcaggg caagaaggtc
     2581 cgcaagccca gcagcaaagg cctggcctgt ggaagcaagg aggccaagga cctcaaggca
     2641 cggaggaaga agtcccagga cggcaagggc tgcctgctgg acagctccgg catgctctcg
     2701 cccgtggact ccctggagtc accccatggc tacctgtcag acgtggcctc gccgccactg
     2761 ctgccctccc cgttccagca gtctccgtcc gtgcccctca accacctgcc tgggatgccc
     2821 gacacccacc tgggcatcgg gcacctgaac gtggcggcca agcccgagat ggcggcgctg
     2881 ggtgggggcg gccggctggc ctttgagact ggcccacctc gtctctccca cctgcctgtg
     2941 gcctctggca ccagcaccgt cctgggctcc agcagcggag gggccctgaa tttcactgtg
     3001 ggcgggtcca ccagtttgaa tggtcaatgc gagtggctgt cccggctgca gagcggcatg
     3061 gtgccgaacc aatacaaccc tctgcggggg agtgtggcac caggccccct gagcacacag
     3121 gccccctccc tgcagcatgg catggtaggc ccgctgcaca gtagccttgc tgccagcgcc
     3181 ctgtcccaga tgatgagcta ccagggcctg cccagcaccc ggctggccac ccagcctcac
     3241 ctggtgcaga cccagcaggt gcagccacaa aacttacaga tgcagcagca gaacctgcag
     3301 ccagcaaaca tccagcagca gcaaagcctg cagccgccac caccaccacc acagccgcac
     3361 cttggcgtga gctcagcagc cagcggccac ctgggccgga gcttcctgag tggagagccg
     3421 agccaggcag acgtgcagcc actgggcccc agcagcctgg cggtgcacac tattctgccc
     3481 caggagagcc ccgccctgcc cacgtcgctg ccatcctcgc tggtcccacc cgtgaccgca
     3541 gcccagttcc tgacgccccc ctcgcagcac agctactcct cgcctgtgga caacaccccc
     3601 agccaccagc tacaggtgcc tgagcacccc ttcctcaccc cgtcccctga gtcccctgac
     3661 cagtggtcca gctcgtcccc gcattccaac gtctccgact ggtccgaggg cgtctccagc
     3721 cctcccacca gcatgcagtc ccagatcgcc cgcattccgg aggccttcaa ggattacaag
     3781 gacgacgatg acaagtaaga attcgccagc acagtggtcg accaattccg gttattttcc
     3841 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     3901 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     3961 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     4021 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     4081 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     4141 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     4201 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     4261 aggttaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     4321 gatgataata ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     4381 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     4441 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     4501 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     4561 gtgcttgaca ttggggaatt tagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     4621 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     4681 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     4741 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     4801 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     4861 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     4921 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     4981 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     5041 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     5101 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     5161 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     5221 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     5281 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     5341 ccgagggcaa aggaatagag tagatgccga ccgggatcta tcgataaaat aaaagatttt
     5401 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     5461 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     5521 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     5581 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     5641 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     5701 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     5761 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     5821 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     5881 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     5941 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     6001 cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta
     6061 tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6121 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6181 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6241 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6301 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6361 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6421 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6481 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6541 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6601 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6661 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6721 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6781 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6841 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6901 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6961 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc
     7021 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7081 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7141 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7201 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7261 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7321 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7381 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7441 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7501 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7561 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7621 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7681 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7741 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7801 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7861 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7921 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7981 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8041 cgaaaagtgc cac
//
