LOCUS       sequence-329773-        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"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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"
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2223..3401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3349..3369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3414..3443
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5115..5576
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5171..5188)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5398..5417
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    6038..6626
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6091..6111)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6509..6520)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6774..6954
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7001..7019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7087..7191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7192..7260,7261..8052)
                     /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(7410..7429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8223..8811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8965..8982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9222..9243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9258..9263,9264..9281,9282..9288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9296..9312
                     /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     9301..9323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9320..9336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9320..9336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9386..9405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagcttcagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccgc atggaacaaa
     3421 agttgatttc tgaagaagat ttgttaagaa agagaggatc cactgaggac cacaaaatgg
     3481 tagaagcctt acgtgtttgg gcatctactc atatgtcacc gtcttggaca ttactaaaat
     3541 tgtgtgatgt tcagccaatg cagtattttg acctgacttg tcagctcttg ggcaaagcag
     3601 aagtggacgg agcatcattt cttctaaagg tatgggatgg caccaggaca ccatttccat
     3661 cttggagagt cttaatacaa gaccttgttc ttgaaggtga tttaagtcac atccatcggc
     3721 tacaaaatct gacaatagac attttagtct acgataacca tgttcatgtg gcaagatctc
     3781 tgaaggttgg aagctttctt agaatctata gccttcatac caaacttcaa tcaatgaatt
     3841 cagagaatca gacaatgtta agtttagagt ttcatcttca tggaggtacc agttacggtc
     3901 ggggaatcag ggtcttgcca gaaagtaact ctgatgtgga tcaactgaaa aaggatttag
     3961 aatctgcaaa tttgacagcc aatcagcatt cagatgttat ctgtcaatca gaacctgacg
     4021 acagctttcc aagctctgga tcagtatcat tatacgaggt agaaagatgt caacagctat
     4081 ctgctacaat acttacagat catcagtatt tggagaggac accactatgt gccattttga
     4141 aacaaaaagc tcctcaacaa taccgcatcc gagcaaaatt gaggtcatat aagcccagaa
     4201 gactatttca gtctgttaaa cttcattgcc ctaaatgtca tttgctgcaa gaagttccac
     4261 atgagggcga tttggatata atttttcagg atggtgcaac taaaacccca gatgtcaagc
     4321 tacaaaatac atcattatat gattcaaaaa tctggaccac taaaaatcaa aaaggacgaa
     4381 aagtagcagt tcattttgtg aaaaataatg gtattctccc gctttcaaat gaatgtctac
     4441 ttttgataga aggaggtaca ctcagtgaaa tttgcaaact ctcgaacaag tttaatagtg
     4501 taattcctgt gagatctggc cacgaagacc tggaactttt ggacctttca gcaccatttc
     4561 ttatacaagg aacaatacat cactatggat gtaaacagtg ttctagtttg agatccatac
     4621 aaaatctaaa ttccctggtt gataaaacat cgtggattcc ttcttctgtg gcagaagcac
     4681 tgggtattgt acccctccaa tatgtgtttg ttatgacctt tacacttgat gatggaacag
     4741 gagtactaga agcctatctc atggattctg acaaattctt ccagattcca gcatcagaag
     4801 ttctgatgga tgatgacctt cagaaaagtg tggatatgat catggatatg ttttgtcctc
     4861 caggaataaa aattgatgca tatccgtggt tggaatgctt catcaagtca tacaatgtca
     4921 caaatggaac agataatcaa atttgctatc agatttttga caccacagtt gcagaagatg
     4981 taatctaatt aattaagttt aaacggatcc cgccggcggc tagcgttaac acgcgttgta
     5041 cattttacgt atctagagct cgaggtcgac caattccggt tattttccac catattgccg
     5101 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     5161 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     5221 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     5281 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     5341 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     5401 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     5461 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac
     5521 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatacc
     5581 atgaaaacat ttaacatttc tcaacaggat ctagaattag tagaagtagc gacagagaag
     5641 attacaatgc tttatgagga taataaacat catgtgggag cggcaattcg tacgaaaaca
     5701 ggagaaatca tttcggcagt acatattgaa gcgtatatag gacgagtaac tgtttgtgca
     5761 gaagccattg cgattggtag tgcagtttcg aatggacaaa aggattttga cacgattgta
     5821 gctgttagac acccttattc tgacgaagta gatagaagta ttcgagtggt aagtccttgt
     5881 ggtatgtgta gggagttgat ttcagactat gcaccagatt gttttgtgtt aatagaaatg
     5941 aatggcaagt tagtcaaaac tacgattgaa gaactcattc cactcaaata tacccgaaat
     6001 taaaagtttt accataccaa gctcatcgat agatcctaat caacctctgg attacaaaat
     6061 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6121 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6181 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     6241 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6301 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6361 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6421 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6481 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6541 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6601 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     6661 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6721 aacgaagaca agatcacctg caggacaggc gcgccctgct ttttgcttgt actgggtctc
     6781 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6841 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6901 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcacccggg
     6961 cgattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat acgcctattt
     7021 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7201 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7261 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7321 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7381 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     7441 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     7501 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7561 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7621 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7681 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     7741 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7801 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7861 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     7921 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7981 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8041 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     8341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     8401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8881 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8941 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     9001 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct
     9061 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     9121 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gtggcacgac
     9181 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     9241 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     9301 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt cacaaataaa
     9361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     9421 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
     9481 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
     9541 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
