LOCUS       sequence-466041-        8041 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Myc-His-BirA(R118G)-TRF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8041)
  AUTHORS   Garcia-Exposito L, Bournique E, Bergoglio V, Bose A, 
            Barroso-Gonzalez J, Zhang S, Roncaioli JL, Lee M, Wallace CT, 
            Watkins SC, Opresko PL, Hoffmann JS, O'Sullivan RJ
  TITLE     Proteomic Profiling Reveals a Specific Role for Translesion DNA 
            Polymerase eta in the Alternative Lengthening of Telomeres.
  JOURNAL   Cell Rep. 2016 Nov 8;17(7):1858-1871. doi: 
            10.1016/j.celrep.2016.10.048.
  PUBMED    27829156
REFERENCE   2  (bases 1 to 8041)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2016.10"; journalName: "Cell Rep"; date: 
            "2016-11-8- 8"; volume: "17"; issue: "7"; pages: "1858-1871"
COMMENT     Sequence Label: pLPC-Myc-BirA-TRF1
FEATURES             Location/Qualifiers
     source          1..8041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             52..521
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    586..785
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             986..1402
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1332..1354
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1370..1386
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1420..2019
                     /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(1420..1439)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1916..1936
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2108..2127)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2299..2678
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2679..2882
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2833..2853
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2927..2946
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2927..2945
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2977..2986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2986..3015
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3034..3051
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(3052..3399,3400..3402,3403..4011)
                     /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             5579..6048
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6178..6195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6349..6937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6429..6448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7097..7888,7889..7957))
                     /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     7720..7739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7958..8041,1..21))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg
       61 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
      121 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
      181 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
      241 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
      301 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
      361 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
      421 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
      481 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
      541 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac
      601 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta
      661 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
      721 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
      781 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
      841 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
      901 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
      961 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     1021 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     1081 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     1141 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     1201 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     1261 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     1321 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     1381 cctcactcct tctctaggcg ccggccggat cagcttacca tgaccgagta caagcccacg
     1441 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     1501 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     1561 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     1621 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     1681 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     1741 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     1801 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     1861 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     1921 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     1981 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     2041 cgcccgaccg aaaggagcgc acgaccccat gcatcggatc tcccgatccc ctatggtgca
     2101 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     2161 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     2221 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     2281 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     2341 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2401 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2461 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2521 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2581 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2641 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2701 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2761 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2821 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2881 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     2941 atagggagac ccaagcttgg taccgagctc ggatcagcca ccatggaaca aaagttgatt
     3001 tctgaagaag atttgttaag aaagagagga tcccatcatc accatcacca caaggacaac
     3061 accgtgcccc tgaagctgat cgccctgctg gccaatggcg agttccactc tggcgagcag
     3121 ctgggcgaga cactgggcat gtctagagcc gccatcaaca agcacatcca gaccctgaga
     3181 gactggggcg tggacgtgtt caccgtgcct ggcaagggct acagcctgcc cgagcctatc
     3241 cagctgctga acgccaagca gatcctgggc cagctggatg gcggatctgt ggctgtgctg
     3301 cccgtgatcg acagcaccaa ccagtacctg ctggaccgga tcggcgagct gaaaagcggc
     3361 gacgcctgta tcgccgagta tcagcaggct ggcagaggcg gcaggggcag aaagtggttc
     3421 tctcccttcg gcgccaatct gtacctgagc atgttctggc ggctggaaca gggacctgcc
     3481 gccgctattg gactgagcct cgtgatcggc atcgtgatgg ccgaggtgct gagaaagctg
     3541 ggcgccgaca aagtgcgcgt gaagtggccc aacgacctgt atctgcagga ccggaagctg
     3601 gccggcatcc tggtggaact gacaggcaaa acaggcgacg ctgcccagat cgtgattggc
     3661 gctggcatca atatggccat gcggcgggtg gaagagtccg tcgtgaatca gggctggatc
     3721 accctgcagg aagccggcat caacctggac agaaacaccc tggccgccat gctgatcaga
     3781 gagctgagag ccgccctgga actgttcgag caggaaggcc tggcccccta cctgtccaga
     3841 tgggagaagc tggacaactt catcaacaga cccgtgaaac tgatcatcgg cgacaaagag
     3901 atcttcggca tcagccgggg catcgacaag cagggcgctc tgctgctgga acaggacggc
     3961 atcatcaagc cctggatggg cggcgagatc agcctgagat ctgccgagaa aggcggaggc
     4021 agtggcgggg gttcaggatc cgcggaggat gtttcctcag cggccccgag cccgcggggc
     4081 tgtgcggatg gtagggatgc cgaccctact gaggagcaga tggcagaaac agagagaaac
     4141 gacgaggagc agttcgaatg ccaggaactg ctcgagtgcc aggtgcaggt gggggccccc
     4201 gaggaggagg aggaggagga ggaggacgcg ggcctggtgg ccgaggccga ggccgtggct
     4261 gccggctgga tgctcgattt cctctgcctc tctctttgcc gagctttccg cgacggccgc
     4321 tccgaggact tccgcaggac ccgcaacagc gcagaggcta ttattcatgg actatccagt
     4381 ctaacagctt gccagttgag aacgatatac atatgtcagt ttttgacaag aattgcagca
     4441 ggaaaaaccc ttgatgcaca gtttgaaaat gatgaacgaa ttacaccctt ggaatcagcc
     4501 ctgatgattt ggggttcaat tgaaaaggaa catgacaaac ttcatgaaga aatacagaat
     4561 ttaattaaaa ttcaggctat agctgtttgt atggaaaatg gcaactttaa agaagcagaa
     4621 gaagtctttg aaagaatatt tggtgatcca aattctcata tgcctttcaa aagcaaattg
     4681 cttatgataa tctctcagaa agatacattt cattcctttt ttcaacactt cagctacaac
     4741 cacatgatgg agaaaattaa gagttatgtg aattatgtgc taagtgaaaa atcatcaacc
     4801 tttctaatga aggcagcggc aaaagtagta gaaagcaaaa ggacaagaac aataacttct
     4861 caagataaac ctagtggtaa tgatgttgaa atggaaactg aagctaattt ggatacaaga
     4921 aaaagtgtta gtgacaaaca gtctgcggta actgaatcct cagagggtac agtatcctta
     4981 ttgaggtctc acaagaatct tttcttatct aagttgcaac atggaaccca gcaacaagac
     5041 cttaataaga aagaaagaag agtaggaact cctcaaagta caaaaaagaa aaaagaaagc
     5101 agaagagcca ctgaaagcag aatacctgtt tcaaagagtc agccggtaac tcctgaaaaa
     5161 catcgagcta gaaaaagaca ggcatggctt tgggaagaag acaagaattt gagatctggc
     5221 gtgaggaaat atggagaggg aaactggtct aaaatactgt tgcattataa attcaacaac
     5281 cggacaagtg tcatgttaaa agacagatgg aggaccatga agaaactaaa actgatttcc
     5341 tcagacagcg aagactgaga attctgcaga tatccatcac actggcggcc gctcgagcat
     5401 gcatctagag ggccgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     5461 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     5521 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
     5581 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     5641 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     5701 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
     5761 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     5821 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     5881 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     5941 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     6001 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt
     6061 atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta
     6121 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6181 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6241 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6301 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6361 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6421 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6481 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6541 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6601 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6661 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6721 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6781 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6841 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6901 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6961 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7021 aaaaaggatc ttcacctaga tccttttgcg gccgcaaatc aatctaaagt atatatgagt
     7081 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7261 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7621 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7681 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7921 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7981 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8041 a
//
