LOCUS       sequence-329225-        8596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express BirA-hPOT1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8596)
  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 8596)
  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
FEATURES             Location/Qualifiers
     source          1..8596
                     /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             6083..6552
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6682..6699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6853..7441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6933..6952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7601..8392,8393..8461))
                     /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     8224..8243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8462..8566)
                     /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 ggatccttgg ttccagcaac aaattatata
     4021 tatacacccc tgaatcaact taagggtggt acaattgtca atgtctatgg tgttgtgaag
     4081 ttctttaagc ccccatatct aagcaaagga actgattatt gctcagttgt aactattgtg
     4141 gaccagacaa atgtaaaact aacttgcctg ctctttagtg gaaactatga agcccttcca
     4201 ataatttata aaaatggaga tattgttcgc tttcacaggc tgaagattca agtatataaa
     4261 aaggagactc agggtatcac cagctctggc tttgcatctt tgacgtttga gggaactttg
     4321 ggagccccta tcatacctcg cacttcaagc aagtatttta acttcactac tgaggaccac
     4381 aaaatggtag aagccttacg tgtttgggca tctactcata tgtcaccgtc ttggacatta
     4441 ctaaaattgt gtgatgttca gccaatgcag tattttgacc tgacttgtca gctcttgggc
     4501 aaagcagaag tggacggagc atcatttctt ctaaaggtat gggatggcac caggacacca
     4561 tttccatctt ggagagtctt aatacaagac cttgttcttg aaggtgattt aagtcacatc
     4621 catcggctac aaaatctgac aatagacatt ttagtctacg ataaccatgt tcatgtggca
     4681 agatctctga aggttggaag ctttcttaga atctatagcc ttcataccaa acttcaatca
     4741 atgaattcag agaatcagac aatgttaagt ttagagtttc atcttcatgg aggtaccagt
     4801 tacggtcggg gaatcagggt cttgccagaa agtaactctg atgtggatca actgaaaaag
     4861 gatttagaat ctgcaaattt gacagccaat cagcattcag atgttatctg tcaatcagaa
     4921 cctgacgaca gctttccaag ctctggatca gtatcattat acgaggtaga aagatgtcaa
     4981 cagctatctg ctacaatact tacagatcat cagtatttgg agaggacacc actatgtgcc
     5041 attttgaaac aaaaagctcc tcaacaatac cgcatccgag caaaattgag gtcatataag
     5101 cccagaagac tatttcagtc tgttaaactt cattgcccta aatgtcattt gctgcaagaa
     5161 gttccacatg agggcgattt ggatataatt tttcaggatg gtgcaactaa aaccccagat
     5221 gtcaagctac aaaatacatc attatatgat tcaaaaatct ggaccactaa aaatcaaaaa
     5281 ggacgaaaag tagcagttca ttttgtgaaa aataatggta ttctcccgct ttcaaatgaa
     5341 tgtctacttt tgatagaagg aggtacactc agtgaaattt gcaaactctc gaacaagttt
     5401 aatagtgtaa ttcctgtgag atctggccac gaagacctgg aacttttgga cctttcagca
     5461 ccatttctta tacaaggaac aatacatcac tatggatgta aacagtgttc tagtttgaga
     5521 tccatacaaa atctaaattc cctggttgat aaaacatcgt ggattccttc ttctgtggca
     5581 gaagcactgg gtattgtacc cctccaatat gtgtttgtta tgacctttac acttgatgat
     5641 ggaacaggag tactagaagc ctatctcatg gattctgaca aattcttcca gattccagca
     5701 tcagaagttc tgatggatga tgaccttcag aaaagtgtgg atatgatcat ggatatgttt
     5761 tgtcctccag gaataaaaat tgatgcatat ccgtggttgg aatgcttcat caagtcatac
     5821 aatgtcacaa atggaacaga taatcaaatt tgctatcaga tttttgacac cacagttgca
     5881 gaagatgtaa tctaactcga gcatgcatct agagggccga taaaataaaa gattttattt
     5941 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     6001 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca
     6061 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     6121 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
     6181 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     6241 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     6301 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     6361 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct
     6421 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     6481 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     6541 gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta
     6601 cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6661 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6721 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6781 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6841 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6901 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6961 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7021 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7081 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7141 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7201 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7261 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7321 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7381 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7441 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7501 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccgca
     7561 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7681 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7741 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8461 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8521 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8581 aagtgccacc tgcagc
//
