LOCUS       sequence-333514-        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             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"
     regulatory      1362..1371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1371..1400
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3368..3829
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3424..3441)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3651..3670
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3834..4859
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3834..4859
                     /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             5048..5517
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5647..5664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5818..6406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5898..5917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6566..7357,7358..7426))
                     /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     7189..7208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7427..7531)
                     /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 agccaccatg gaacaaaagt
     1381 tgatttctga agaagatttg ttaagaaaga gaggatcctt ggttccagca acaaattata
     1441 tatatacacc cctgaatcaa cttaagggtg gtacaattgt caatgtctat ggtgttgtga
     1501 agttctttaa gcccccatat ctaagcaaag gaactgatta ttgctcagtt gtaactattg
     1561 tggaccagac aaatgtaaaa ctaacttgcc tgctctttag tggaaactat gaagcccttc
     1621 caataattta taaaaatgga gatattgttc gctttcacag gctgaagatt caagtatatg
     1681 agaaggagac tcagggtatc accagctctg gctttgcatc tttgacgttt gagggaactt
     1741 tgggagcccc tatcatacct cgcacttcaa gcaagtattt taacttcact actgaggacc
     1801 acaaaatggt agaagcctta cgtgtttggg catctactca tatgtcaccg tcttggacat
     1861 tactaaaatt gtgtgatgtt cagccaatgc agtattttga cctgacttgt cagctcttgg
     1921 gcaaagcaga agtggacgga gcatcatttc ttctaaaggt atgggatggc accaggacac
     1981 catttccatc ttggagagtc ttaatacaag accttgttct tgaaggtgat ttaagtcaca
     2041 tccatcggct acaaaatctg acaatagaca ttttagtcta cgataaccat gttcatgtgg
     2101 caagatctct gaaggttgga agctttctta gaatctatag ccttcatacc aaacttcaat
     2161 caatgaattc agagaatcag acaatgttaa gtttagagtt tcatcttcat ggaggtacca
     2221 gttacggtcg gggaatcagg gtcttgccag aaagtaactc tgatgtggat caactgaaaa
     2281 aggatttaga atctgcaaat ttgacagcca atcagcattc agatgttatc tgtcaatcag
     2341 aacctgacga cagctttcca agctctggat cagtatcatt atacgaggta gaaagatgtc
     2401 aacagctatc tgctacaata cttacagatc atcagtattt ggagaggaca ccactatgtg
     2461 ccattttgaa acaaaaagct cctcaacaat accgcatccg agcaaaattg aggtcatata
     2521 agcccagaag actatttcag tctgttaaac ttcattgccc taaatgtcat ttgctgcaag
     2581 aagttccaca tgagggcgat ttggatataa tttttcagga tggtgcaact aaaaccccag
     2641 atgtcaagct acaaaataca tcattatatg attcaaaaat ctggaccact aaaaatcaaa
     2701 aaggacgaaa agtagcagtt cattttgtga aaaataatgg tattctcccg ctttcaaatg
     2761 aatgtctact tttgatagaa ggaggtacac tcagtgaaat ttgcaaactc tcgaacaagt
     2821 ttaatagtgt aattcctgtg agatctggcc acgaagacct ggaacttttg gacctttcag
     2881 caccatttct tatacaagga acaatacatc actatggatg taaacagtgt tctagtttga
     2941 gatccataca aaatctaaat tccctggttg ataaaacatc gtggattcct tcttctgtgg
     3001 cagaagcact gggtattgta cccctccaat atgtgtttgt tatgaccttt acacttgatg
     3061 atggaacagg agtactagaa gcctatctca tggattctga caaattcttc cagattccag
     3121 catcagaagt tctgatggat gatgaccttc agaaaagtgt ggatatgatc atggatatgt
     3181 tttgtcctcc aggaataaaa attgatgcat atccgtggtt ggaatgcttc atcaagtcat
     3241 acaatgtcac aaatggaaca gataatcaaa tttgctatca gatttttgac accacagttg
     3301 cagaagatgt aatctaactc gaccaattcc ggttattttc caccatattg ccgtcttttg
     3361 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     3421 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     3481 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     3541 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     3601 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     3661 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     3721 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg
     3781 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat accatgaaaa
     3841 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     3901 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     3961 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     4021 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
     4081 ttagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     4141 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     4201 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     4261 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     4321 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     4381 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     4441 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     4501 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     4561 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     4621 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     4681 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     4741 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     4801 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataga
     4861 gtagatgccg accgggatct atcgataaaa taaaagattt tatttagtct ccagaaaaag
     4921 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     4981 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat
     5041 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     5101 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     5161 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
     5221 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     5281 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     5341 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
     5401 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     5461 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat
     5521 gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata
     5581 gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6481 gagattatca aaaaggatct tcacctagat ccttttgcgg ccgcaaatca atctaaagta
     6541 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6601 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6661 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6721 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6781 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6841 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6901 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6961 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7021 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7081 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7141 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7201 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7261 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7321 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7381 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7441 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7501 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag
     7561 c
//
