LOCUS       sequence-324922-        7561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hPOT1 K90E in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7561)
  AUTHORS   Pinzaru AM, Kareh M, Lamm N, Lazzerini-Denchi E, Cesare AJ, Sfeir A
  TITLE     Replication stress conferred by POT1 dysfunction promotes telomere 
            relocalization to the nuclear pore.
  JOURNAL   Genes Dev. 2020 Dec 1;34(23-24):1619-1636. doi: 
            10.1101/gad.337287.120. Epub 2020 Oct 29.
  PUBMED    33122293
REFERENCE   2  (bases 1 to 7561)
  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: "Genes 
            Dev."; date: "2020-12-1"; volume: "34(23-24):1619-1636. doi"; pages:
            " 10.1101/gad.337287.120. Epub 2020 Oct 29"
COMMENT     Sequence Label: pWZL N-Myc-hPOT1_K90E-IRES_hygro
FEATURES             Location/Qualifiers
     source          1..7561
                     /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"
     regulatory      1369..1378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1378..1407
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3375..3836
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3431..3448)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3658..3677
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3841..4866
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3841..4866
                     /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             5055..5524
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5654..5671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5825..6413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5905..5924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6573..7364,7365..7433))
                     /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     7196..7215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7434..7538)
                     /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 ccggatcagc caccatggaa
     1381 caaaagttga tttctgaaga agatttgtta agaaagagag gatccttggt tccagcaaca
     1441 aattatatat atacacccct gaatcaactt aagggtggta caattgtcaa tgtctatggt
     1501 gttgtgaagt tctttaagcc cccatatcta agcaaaggaa ctgattattg ctcagttgta
     1561 actattgtgg accagacaaa tgtaaaacta acttgcctgc tctttagtgg aaactatgaa
     1621 gcccttccaa taatttataa aaatggagat attgttcgct ttcacaggct gaagattcaa
     1681 gtatatgaga aggagactca gggtatcacc agctctggct ttgcatcttt gacgtttgag
     1741 ggaactttgg gagcccctat catacctcgc acttcaagca agtattttaa cttcactact
     1801 gaggaccaca aaatggtaga agccttacgt gtttgggcat ctactcatat gtcaccgtct
     1861 tggacattac taaaattgtg tgatgttcag ccaatgcagt attttgacct gacttgtcag
     1921 ctcttgggca aagcagaagt ggacggagca tcatttcttc taaaggtatg ggatggcacc
     1981 aggacaccat ttccatcttg gagagtctta atacaagacc ttgttcttga aggtgattta
     2041 agtcacatcc atcggctaca aaatctgaca atagacattt tagtctacga taaccatgtt
     2101 catgtggcaa gatctctgaa ggttggaagc tttcttagaa tctatagcct tcataccaaa
     2161 cttcaatcaa tgaattcaga gaatcagaca atgttaagtt tagagtttca tcttcatgga
     2221 ggtaccagtt acggtcgggg aatcagggtc ttgccagaaa gtaactctga tgtggatcaa
     2281 ctgaaaaagg atttagaatc tgcaaatttg acagccaatc agcattcaga tgttatctgt
     2341 caatcagaac ctgacgacag ctttccaagc tctggatcag tatcattata cgaggtagaa
     2401 agatgtcaac agctatctgc tacaatactt acagatcatc agtatttgga gaggacacca
     2461 ctatgtgcca ttttgaaaca aaaagctcct caacaatacc gcatccgagc aaaattgagg
     2521 tcatataagc ccagaagact atttcagtct gttaaacttc attgccctaa atgtcatttg
     2581 ctgcaagaag ttccacatga gggcgatttg gatataattt ttcaggatgg tgcaactaaa
     2641 accccagatg tcaagctaca aaatacatca ttatatgatt caaaaatctg gaccactaaa
     2701 aatcaaaaag gacgaaaagt agcagttcat tttgtgaaaa ataatggtat tctcccgctt
     2761 tcaaatgaat gtctactttt gatagaagga ggtacactca gtgaaatttg caaactctcg
     2821 aacaagttta atagtgtaat tcctgtgaga tctggccacg aagacctgga acttttggac
     2881 ctttcagcac catttcttat acaaggaaca atacatcact atggatgtaa acagtgttct
     2941 agtttgagat ccatacaaaa tctaaattcc ctggttgata aaacatcgtg gattccttct
     3001 tctgtggcag aagcactggg tattgtaccc ctccaatatg tgtttgttat gacctttaca
     3061 cttgatgatg gaacaggagt actagaagcc tatctcatgg attctgacaa attcttccag
     3121 attccagcat cagaagttct gatggatgat gaccttcaga aaagtgtgga tatgatcatg
     3181 gatatgtttt gtcctccagg aataaaaatt gatgcatatc cgtggttgga atgcttcatc
     3241 aagtcataca atgtcacaaa tggaacagat aatcaaattt gctatcagat ttttgacacc
     3301 acagttgcag aagatgtaat ctaactcgac caattccggt tattttccac catattgccg
     3361 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     3421 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     3481 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     3541 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     3601 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     3661 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     3721 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac
     3781 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatacc
     3841 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     3901 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     3961 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     4021 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     4081 ggggaattta gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     4141 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     4201 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     4261 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     4321 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     4381 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     4441 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     4501 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     4561 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     4621 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     4681 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     4741 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     4801 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     4861 gaatagagta gatgccgacc gggatctatc gataaaataa aagattttat ttagtctcca
     4921 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     4981 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     5041 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5101 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     5161 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     5221 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     5281 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     5341 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     5401 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     5461 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5521 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     5581 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6241 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg caaatcaatc
     6541 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6601 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6661 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6721 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6781 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6841 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6901 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6961 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7021 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7081 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7141 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7201 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7261 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7321 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7381 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7441 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7501 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7561 c
//
