LOCUS       sequence-331987-        8134 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 8134)
  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 8134)
  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..8134
                     /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             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1280..1302
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3764..3787
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3859..4320
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3915..3932)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4142..4161
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4325..5350
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGAFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             5415..6008
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6209..6226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6380..6968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6460..6479)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7146..7937,7938..8006))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7769..7788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8007..8111)
                     /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 taagctggcc 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 tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     1261 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc gccaccatgc gcaagcgccg
     1381 gcggcagcat ggccagctct ggttccctga gggcttcaaa gtgtctgagg ccagcaagaa
     1441 gaagcggcgg gagcccctcg gcgaggactc cgtgggcctc aagcccctga agaacgcttc
     1501 agacggtgcc ctcatggacg acaaccagaa tgagtggggg gacgaggacc tggagaccaa
     1561 gaagttccgg ttcgaggagc ccgtggttct gcctgacctg gacgaccaga cagaccaccg
     1621 gcagtggact cagcagcacc tggatgccgc tgacctgcgc atgtctgcca tggcccccac
     1681 accgccccag ggtgaggttg acgccgactg catggacgtc aatgtccgcg ggcctgatgg
     1741 cttcaccccg ctcatgatcg cctcctgcag cgggggcggc ctggagacgg gcaacagcga
     1801 ggaagaggag gacgcgccgg ccgtcatctc cgacttcatc taccagggcg ccagcctgca
     1861 caaccagaca gaccgcacgg gcgagaccgc cttgcacctg gccgcccgct actcacgctc
     1921 tgatgccgcc aagcgcctgc tggaggccag cgcagatgcc aacatccagg acaacatggg
     1981 ccgcaccccg ctgcatgcgg ctgtgtctgc cgacgcacaa ggtgtcttcc agatcctgat
     2041 ccggaaccga gccacagacc tggatgcccg catgcatgat ggcacgacgc cactgatcct
     2101 ggctgcccgc ctggccgtgg agggcatgct ggaggacctc atcaactcac acgccgacgt
     2161 caacgccgta gatgacctgg gcaagtccgc cctgcactgg gccgccgccg tgaacaatgt
     2221 ggatgccgca gttgtgctcc tgaagaacgg ggctaacaaa gatatgcaga acaacaggga
     2281 ggagacaccc ctgtttctgg ccgcccggga gggcagctac gagaccgcca aggtgctgct
     2341 ggaccacttt gccaaccggg acatcacgga tcatatggac cgcctgccgc gcgacatcgc
     2401 acaggagcgc atgcatcacg acatcgtgag gctgctggac gagtacaacc tggtgcgcag
     2461 cccgcagctg cacggagccc cgctgggggg cacgcccacc ctgtcgcccc cgctctgctc
     2521 gcccaacggc tacctgggca gcctcaagcc cggcgtgcag ggcaagaagg tccgcaagcc
     2581 cagcagcaaa ggcctggcct gtggaagcaa ggaggccaag gacctcaagg cacggaggaa
     2641 gaagtcccag gacggcaagg gctgcctgct ggacagctcc ggcatgctct cgcccgtgga
     2701 ctccctggag tcaccccatg gctacctgtc agacgtggcc tcgccgccac tgctgccctc
     2761 cccgttccag cagtctccgt ccgtgcccct caaccacctg cctgggatgc ccgacaccca
     2821 cctgggcatc gggcacctga acgtggcggc caagcccgag atggcggcgc tgggtggggg
     2881 cggccggctg gcctttgaga ctggcccacc tcgtctctcc cacctgcctg tggcctctgg
     2941 caccagcacc gtcctgggct ccagcagcgg aggggccctg aatttcactg tgggcgggtc
     3001 caccagtttg aatggtcaat gcgagtggct gtcccggctg cagagcggca tggtgccgaa
     3061 ccaatacaac cctctgcggg ggagtgtggc accaggcccc ctgagcacac aggccccctc
     3121 cctgcagcat ggcatggtag gcccgctgca cagtagcctt gctgccagcg ccctgtccca
     3181 gatgatgagc taccagggcc tgcccagcac ccggctggcc acccagcctc acctggtgca
     3241 gacccagcag gtgcagccac aaaacttaca gatgcagcag cagaacctgc agccagcaaa
     3301 catccagcag cagcaaagcc tgcagccgcc accaccacca ccacagccgc accttggcgt
     3361 gagctcagca gccagcggcc acctgggccg gagcttcctg agtggagagc cgagccaggc
     3421 agacgtgcag ccactgggcc ccagcagcct ggcggtgcac actattctgc cccaggagag
     3481 ccccgccctg cccacgtcgc tgccatcctc gctggtccca cccgtgaccg cagcccagtt
     3541 cctgacgccc ccctcgcagc acagctactc ctcgcctgtg gacaacaccc ccagccacca
     3601 gctacaggtg cctgagcacc ccttcctcac cccgtcccct gagtcccctg accagtggtc
     3661 cagctcgtcc ccgcattcca acgtctccga ctggtccgag ggcgtctcca gccctcccac
     3721 cagcatgcag tcccagatcg cccgcattcc ggaggccttc aaggattaca aggacgacga
     3781 tgacaagtaa gaattcgcca gcacagtggt cgaccaattc cggttatttt ccaccatatt
     3841 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     3901 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     3961 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     4021 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     4081 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     4141 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     4201 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     4261 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     4321 taccatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     4381 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     4441 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     4501 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     4561 cattggggca ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     4621 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     4681 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     4741 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     4801 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     4861 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     4921 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     4981 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     5041 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     5101 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     5161 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     5221 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     5281 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     5341 aaaggaatag gcgggactct ggggttcgca tcgataaaat aaaagatttt atttagtctc
     5401 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     5461 ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca
     5521 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     5581 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5641 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     5701 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     5761 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     5821 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     5881 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     5941 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     6001 ctttcatttc cgacttgtgg tctcgctgcc ttgggagggt ctcctctgag tgattgacta
     6061 cccgtcagcg ggggtcttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     6121 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     6181 ggtttgcgta ttggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6241 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6301 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6361 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6421 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6481 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6541 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6601 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6661 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6721 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6781 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6841 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6901 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6961 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7021 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7081 attaaaaatg aagtttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     7141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7261 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7441 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7741 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     7801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8101 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
