LOCUS       sequence-331505-        8224 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 8224)
  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 8224)
  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..8224
                     /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"
     CDS             1369..1968
                     /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(1369..1388)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1865..1885
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2057..2076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2248..2627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2628..2831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2782..2802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2876..2895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2876..2894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2926..2935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2935..2964
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2983..3006
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(3007..3354,3355..3357,3358..3966)
                     /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"
     LTR             5711..6180
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6310..6327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6481..7069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6561..6580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7229..8020,8021..8089))
                     /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     7852..7871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8090..8194)
                     /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 agcttaccat gaccgagtac
     1381 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     1441 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     1501 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     1561 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     1621 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     1681 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     1741 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     1801 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     1861 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     1921 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     1981 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggatct cccgatcccc
     2041 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     2101 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     2161 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     2221 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     2281 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2341 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2401 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2461 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2521 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2581 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     2641 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     2701 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2761 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2821 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
     2881 cgactcacta tagggagacc caagcttggt accgagctcg gatcagccac catggaacaa
     2941 aagttgattt ctgaagaaga tttgttaaga aagagaggat ccgattacaa ggatgacgac
     3001 gataagaagg acaacaccgt gcccctgaag ctgatcgccc tgctggccaa tggcgagttc
     3061 cactctggcg agcagctggg cgagacactg ggcatgtcta gagccgccat caacaagcac
     3121 atccagaccc tgagagactg gggcgtggac gtgttcaccg tgcctggcaa gggctacagc
     3181 ctgcccgagc ctatccagct gctgaacgcc aagcagatcc tgggccagct ggatggcgga
     3241 tctgtggctg tgctgcccgt gatcgacagc accaaccagt acctgctgga ccggatcggc
     3301 gagctgaaaa gcggcgacgc ctgtatcgcc gagtatcagc aggctggcag aggcggcagg
     3361 ggcagaaagt ggttctctcc cttcggcgcc aatctgtacc tgagcatgtt ctggcggctg
     3421 gaacagggac ctgccgccgc tattggactg agcctcgtga tcggcatcgt gatggccgag
     3481 gtgctgagaa agctgggcgc cgacaaagtg cgcgtgaagt ggcccaacga cctgtatctg
     3541 caggaccgga agctggccgg catcctggtg gaactgacag gcaaaacagg cgacgctgcc
     3601 cagatcgtga ttggcgctgg catcaatatg gccatgcggc gggtggaaga gtccgtcgtg
     3661 aatcagggct ggatcaccct gcaggaagcc ggcatcaacc tggacagaaa caccctggcc
     3721 gccatgctga tcagagagct gagagccgcc ctggaactgt tcgagcagga aggcctggcc
     3781 ccctacctgt ccagatggga gaagctggac aacttcatca acagacccgt gaaactgatc
     3841 atcggcgaca aagagatctt cggcatcagc cggggcatcg acaagcaggg cgctctgctg
     3901 ctggaacagg acggcatcat caagccctgg atgggcggcg agatcagcct gagatctgcc
     3961 gagaaaggcg gaggcagtgg cgggggttca ggatccactg aggaccacaa aatggtagaa
     4021 gccttacgtg tttgggcatc tactcatatg tcaccgtctt ggacattact aaaattgtgt
     4081 gatgttcagc caatgcagta ttttgacctg acttgtcagc tcttgggcaa agcagaagtg
     4141 gacggagcat catttcttct aaaggtatgg gatggcacca ggacaccatt tccatcttgg
     4201 agagtcttaa tacaagacct tgttcttgaa ggtgatttaa gtcacatcca tcggctacaa
     4261 aatctgacaa tagacatttt agtctacgat aaccatgttc atgtggcaag atctctgaag
     4321 gttggaagct ttcttagaat ctatagcctt cataccaaac ttcaatcaat gaattcagag
     4381 aatcagacaa tgttaagttt agagtttcat cttcatggag gtaccagtta cggtcgggga
     4441 atcagggtct tgccagaaag taactctgat gtggatcaac tgaaaaagga tttagaatct
     4501 gcaaatttga cagccaatca gcattcagat gttatctgtc aatcagaacc tgacgacagc
     4561 tttccaagct ctggatcagt atcattatac gaggtagaaa gatgtcaaca gctatctgct
     4621 acaatactta cagatcatca gtatttggag aggacaccac tatgtgccat tttgaaacaa
     4681 aaagctcctc aacaataccg catccgagca aaattgaggt catataagcc cagaagacta
     4741 tttcagtctg ttaaacttca ttgccctaaa tgtcatttgc tgcaagaagt tccacatgag
     4801 ggcgatttgg atataatttt tcaggatggt gcaactaaaa ccccagatgt caagctacaa
     4861 aatacatcat tatatgattc aaaaatctgg accactaaaa atcaaaaagg acgaaaagta
     4921 gcagttcatt ttgtgaaaaa taatggtatt ctcccgcttt caaatgaatg tctacttttg
     4981 atagaaggag gtacactcag tgaaatttgc aaactctcga acaagtttaa tagtgtaatt
     5041 cctgtgagat ctggccacga agacctggaa cttttggacc tttcagcacc atttcttata
     5101 caaggaacaa tacatcacta tggatgtaaa cagtgttcta gtttgagatc catacaaaat
     5161 ctaaattccc tggttgataa aacatcgtgg attccttctt ctgtggcaga agcactgggt
     5221 attgtacccc tccaatatgt gtttgttatg acctttacac ttgatgatgg aacaggagta
     5281 ctagaagcct atctcatgga ttctgacaaa ttcttccaga ttccagcatc agaagttctg
     5341 atggatgatg accttcagaa aagtgtggat atgatcatgg atatgttttg tcctccagga
     5401 ataaaaattg atgcatatcc gtggttggaa tgcttcatca agtcatacaa tgtcacaaat
     5461 ggaacagata atcaaatttg ctatcagatt tttgacacca cagttgcaga agatgtaatc
     5521 taactcgagc atgcatctag agggccgata aaataaaaga ttttatttag tctccagaaa
     5581 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     5641 tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca
     5701 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     5761 tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     5821 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag
     5881 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     5941 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     6001 cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga
     6061 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     6121 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     6181 catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc
     6241 atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7141 catgagatta tcaaaaagga tcttcaccta gatccttttg cggccgcaaa tcaatctaaa
     7201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8221 cagc
//
