LOCUS       sequence-324919-        8006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express BirA-hPOT1 Delta-OB in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8006)
  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 8006)
  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: pLPC-N-Myc-Flag-BirA-hPOT1-DeltaOB
FEATURES             Location/Qualifiers
     source          1..8006
                     /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     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1367..1966
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1367..1386)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1863..1883
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     promoter        2302..2505
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2456..2476
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2549..2568
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2549..2567
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2599..2608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2608..2637
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2656..2679
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2680..3027,3028..3030,3031..3639)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(6081..6098)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6252..6840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6332..6351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7018..7809,7810..7878))
                     /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     7641..7660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7879..7983)
                     /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 tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatca gcttccatga ccgagtacaa
     1381 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     1441 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     1501 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     1561 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     1621 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     1681 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     1741 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     1801 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     1861 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     1921 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     1981 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgat atcggcctgg ctgaccgccc
     2041 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     2101 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     2161 caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc
     2221 tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta catctacgta
     2281 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag
     2341 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     2401 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa
     2461 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
     2521 gaaccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
     2581 ggtaccgagc tcggatcagc caccatggaa caaaagttga tttctgaaga agatttgtta
     2641 agaaagagag gatccgatta caaggatgac gacgataaga aggacaacac cgtgcccctg
     2701 aagctgatcg ccctgctggc caatggcgag ttccactctg gcgagcagct gggcgagaca
     2761 ctgggcatgt ctagagccgc catcaacaag cacatccaga ccctgagaga ctggggcgtg
     2821 gacgtgttca ccgtgcctgg caagggctac agcctgcccg agcctatcca gctgctgaac
     2881 gccaagcaga tcctgggcca gctggatggc ggatctgtgg ctgtgctgcc cgtgatcgac
     2941 agcaccaacc agtacctgct ggaccggatc ggcgagctga aaagcggcga cgcctgtatc
     3001 gccgagtatc agcaggctgg cagaggcggc aggggcagaa agtggttctc tcccttcggc
     3061 gccaatctgt acctgagcat gttctggcgg ctggaacagg gacctgccgc cgctattgga
     3121 ctgagcctcg tgatcggcat cgtgatggcc gaggtgctga gaaagctggg cgccgacaaa
     3181 gtgcgcgtga agtggcccaa cgacctgtat ctgcaggacc ggaagctggc cggcatcctg
     3241 gtggaactga caggcaaaac aggcgacgct gcccagatcg tgattggcgc tggcatcaat
     3301 atggccatgc ggcgggtgga agagtccgtc gtgaatcagg gctggatcac cctgcaggaa
     3361 gccggcatca acctggacag aaacaccctg gccgccatgc tgatcagaga gctgagagcc
     3421 gccctggaac tgttcgagca ggaaggcctg gccccctacc tgtccagatg ggagaagctg
     3481 gacaacttca tcaacagacc cgtgaaactg atcatcggcg acaaagagat cttcggcatc
     3541 agccggggca tcgacaagca gggcgctctg ctgctggaac aggacggcat catcaagccc
     3601 tggatgggcg gcgagatcag cctgagatct gccgagaaag gatccactga ggaccacaaa
     3661 atggtagaag ccttacgtgt ttgggcatct actcatatgt caccgtcttg gacattacta
     3721 aaattgtgtg atgttcagcc aatgcagtat tttgacctga cttgtcagct cttgggcaaa
     3781 gcagaagtgg acggagcatc atttcttcta aaggtatggg atggcaccag gacaccattt
     3841 ccatcttgga gagtcttaat acaagacctt gttcttgaag gtgatttaag tcacatccat
     3901 cggctacaaa atctgacaat agacatttta gtctacgata accatgttca tgtggcaaga
     3961 tctctgaagg ttggaagctt tcttagaatc tatagccttc ataccaaact tcaatcaatg
     4021 aattcagaga atcagacaat gttaagttta gagtttcatc ttcatggagg taccagttac
     4081 ggtcggggaa tcagggtctt gccagaaagt aactctgatg tggatcaact gaaaaaggat
     4141 ttagaatctg caaatttgac agccaatcag cattcagatg ttatctgtca atcagaacct
     4201 gacgacagct ttccaagctc tggatcagta tcattatacg aggtagaaag atgtcaacag
     4261 ctatctgcta caatacttac agatcatcag tatttggaga ggacaccact atgtgccatt
     4321 ttgaaacaaa aagctcctca acaataccgc atccgagcaa aattgaggtc atataagccc
     4381 agaagactat ttcagtctgt taaacttcat tgccctaaat gtcatttgct gcaagaagtt
     4441 ccacatgagg gcgatttgga tataattttt caggatggtg caactaaaac cccagatgtc
     4501 aagctacaaa atacatcatt atatgattca aaaatctgga ccactaaaaa tcaaaaagga
     4561 cgaaaagtag cagttcattt tgtgaaaaat aatggtattc tcccgctttc aaatgaatgt
     4621 ctacttttga tagaaggagg tacactcagt gaaatttgca aactctcgaa caagtttaat
     4681 agtgtaattc ctgtgagatc tggccacgaa gacctggaac ttttggacct ttcagcacca
     4741 tttcttatac aaggaacaat acatcactat ggatgtaaac agtgttctag tttgagatcc
     4801 atacaaaatc taaattccct ggttgataaa acatcgtgga ttccttcttc tgtggcagaa
     4861 gcactgggta ttgtacccct ccaatatgtg tttgttatga cctttacact tgatgatgga
     4921 acaggagtac tagaagccta tctcatggat tctgacaaat tcttccagat tccagcatca
     4981 gaagttctga tggatgatga ccttcagaaa agtgtggata tgatcatgga tatgttttgt
     5041 cctccaggaa taaaaattga tgcatatccg tggttggaat gcttcatcaa gtcatacaat
     5101 gtcacaaatg gaacagataa tcaaatttgc tatcagattt ttgacaccac agttgcagaa
     5161 gatgtaatct aactcgagca tgcatctaga gggccgataa aataaaagat tttatttagt
     5221 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     5281 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     5341 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     5401 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     5461 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     5521 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     5581 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     5641 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     5701 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     5761 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagctcg
     5821 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
     5881 tccgacttgt ggtctcgctg ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5941 cgggggtctt cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta
     6001 cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6061 tattggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6121 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6181 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6241 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6301 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6361 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6421 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6481 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6541 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6601 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6661 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6721 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6781 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6841 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6901 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6961 tgaagtttgc ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7081 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7141 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7201 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7441 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7621 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7681 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7861 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7921 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7981 gcgcacattt ccccgaaaag tgccac
//
