LOCUS       sequence-324753-        9596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express hPOT1 Delta-OB in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9596)
  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 9596)
  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: pHAGE2-EF1a-MCS-dOB-hPOT1_IRES-blast
FEATURES             Location/Qualifiers
     source          1..9596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    19..136
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        184..1362
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          415..1353
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1310..1330
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1375..1404
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    3076..3537
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3132..3149)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3359..3378
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3999..4587
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4052..4072)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4470..4481)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4735..4915
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4962..4980)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5048..5152
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5153..5221,5222..6013)
                     /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     complement(5371..5390)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6184..6772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6926..6943
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7183..7204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7219..7224,7225..7242,7243..7249)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7257..7273
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7262..7284
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7281..7297
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7281..7297
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7347..7366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             7558..8191
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8238..8363
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8860..9093
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9278..9322
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
       61 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      121 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggactagtcg
      181 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
      241 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
      301 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
      361 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
      421 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
      481 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      541 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      601 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      661 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      721 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      781 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      841 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      901 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      961 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     1021 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     1081 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     1141 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1201 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1261 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1321 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaagcggccg catggaacaa
     1381 aagttgattt ctgaagaaga tttgttaaga aagagaggat ccactgagga ccacaaaatg
     1441 gtagaagcct tacgtgtttg ggcatctact catatgtcac cgtcttggac attactaaaa
     1501 ttgtgtgatg ttcagccaat gcagtatttt gacctgactt gtcagctctt gggcaaagca
     1561 gaagtggacg gagcatcatt tcttctaaag gtatgggatg gcaccaggac accatttcca
     1621 tcttggagag tcttaataca agaccttgtt cttgaaggtg atttaagtca catccatcgg
     1681 ctacaaaatc tgacaataga cattttagtc tacgataacc atgttcatgt ggcaagatct
     1741 ctgaaggttg gaagctttct tagaatctat agccttcata ccaaacttca atcaatgaat
     1801 tcagagaatc agacaatgtt aagtttagag tttcatcttc atggaggtac cagttacggt
     1861 cggggaatca gggtcttgcc agaaagtaac tctgatgtgg atcaactgaa aaaggattta
     1921 gaatctgcaa atttgacagc caatcagcat tcagatgtta tctgtcaatc agaacctgac
     1981 gacagctttc caagctctgg atcagtatca ttatacgagg tagaaagatg tcaacagcta
     2041 tctgctacaa tacttacaga tcatcagtat ttggagagga caccactatg tgccattttg
     2101 aaacaaaaag ctcctcaaca ataccgcatc cgagcaaaat tgaggtcata taagcccaga
     2161 agactatttc agtctgttaa acttcattgc cctaaatgtc atttgctgca agaagttcca
     2221 catgagggcg atttggatat aatttttcag gatggtgcaa ctaaaacccc agatgtcaag
     2281 ctacaaaata catcattata tgattcaaaa atctggacca ctaaaaatca aaaaggacga
     2341 aaagtagcag ttcattttgt gaaaaataat ggtattctcc cgctttcaaa tgaatgtcta
     2401 cttttgatag aaggaggtac actcagtgaa atttgcaaac tctcgaacaa gtttaatagt
     2461 gtaattcctg tgagatctgg ccacgaagac ctggaacttt tggacctttc agcaccattt
     2521 cttatacaag gaacaataca tcactatgga tgtaaacagt gttctagttt gagatccata
     2581 caaaatctaa attccctggt tgataaaaca tcgtggattc cttcttctgt ggcagaagca
     2641 ctgggtattg tacccctcca atatgtgttt gttatgacct ttacacttga tgatggaaca
     2701 ggagtactag aagcctatct catggattct gacaaattct tccagattcc agcatcagaa
     2761 gttctgatgg atgatgacct tcagaaaagt gtggatatga tcatggatat gttttgtcct
     2821 ccaggaataa aaattgatgc atatccgtgg ttggaatgct tcatcaagtc atacaatgtc
     2881 acaaatggaa cagataatca aatttgctat cagatttttg acaccacagt tgcagaagat
     2941 gtaatctaat taattaagtt taaacggatc ccgccggcgg ctagcgttaa cacgcgttgt
     3001 acattttacg tatctagagc tcgaggtcga ccaattccgg ttattttcca ccatattgcc
     3061 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     3121 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     3181 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     3241 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     3301 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     3361 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     3421 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     3481 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatac
     3541 catgaaaaca tttaacattt ctcaacagga tctagaatta gtagaagtag cgacagagaa
     3601 gattacaatg ctttatgagg ataataaaca tcatgtggga gcggcaattc gtacgaaaac
     3661 aggagaaatc atttcggcag tacatattga agcgtatata ggacgagtaa ctgtttgtgc
     3721 agaagccatt gcgattggta gtgcagtttc gaatggacaa aaggattttg acacgattgt
     3781 agctgttaga cacccttatt ctgacgaagt agatagaagt attcgagtgg taagtccttg
     3841 tggtatgtgt agggagttga tttcagacta tgcaccagat tgttttgtgt taatagaaat
     3901 gaatggcaag ttagtcaaaa ctacgattga agaactcatt ccactcaaat atacccgaaa
     3961 ttaaaagttt taccatacca agctcatcga tagatcctaa tcaacctctg gattacaaaa
     4021 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     4081 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     4141 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     4201 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     4261 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     4321 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     4381 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     4441 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     4501 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     4561 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca
     4621 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     4681 caacgaagac aagatcacct gcaggacagg cgcgccctgc tttttgcttg tactgggtct
     4741 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     4801 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     4861 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcacccgg
     4921 gcgattaagg aaagggctag atcattcttg aagacgaaag ggcctcgtga tacgcctatt
     4981 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5041 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5101 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5161 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5221 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5281 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5341 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     5401 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5461 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5521 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5581 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5641 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5701 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5761 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5821 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5881 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5941 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6001 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6061 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6121 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6181 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6241 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6301 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6361 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6421 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6481 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6541 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6601 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6661 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6721 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6781 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6841 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6901 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     6961 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagca agctcatggc
     7021 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     7081 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc cgtggcacga
     7141 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7201 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     7261 gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt tcacaaataa
     7321 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7381 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
     7441 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
     7501 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagttgg
     7561 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
     7621 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
     7681 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
     7741 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     7801 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
     7861 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
     7921 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
     7981 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
     8041 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     8101 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     8161 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     8221 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     8281 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     8341 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     8401 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     8461 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     8521 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     8581 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     8641 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     8701 cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     8761 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     8821 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     8881 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     8941 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     9001 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     9061 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     9121 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     9181 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     9241 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     9301 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     9361 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     9421 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     9481 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     9541 agacagatcc attcgattag tgaacggatc tcgacggtat cgccgaattc acaaat
//
