LOCUS       sequence-466042-        6749 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Myc-His-BirA(R118G).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6749)
  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 6749)
  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
FEATURES             Location/Qualifiers
     source          1..6749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        80..184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(185..253,254..1045)
                     /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     complement(403..422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1205..1793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1694..1713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1947..1964
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             2094..2563
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(2839..3447,3448..3450,3451..3798))
                     /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"
     CDS             complement(3799..3816)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3835..3864)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3864..3873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3904..3923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3905..3923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3968..4171)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3997..4017)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4172..4551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4723..4742
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(4831..5430)
                     /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(4914..4934)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5411..5430
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(5448..5864)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5464..5480)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5496..5518)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    6065..6264
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             join(6329..6749,1..49)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagg ctgcaggtgg
       61 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      121 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      181 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      241 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      301 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      361 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      421 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      481 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      541 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      601 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      661 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      721 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      781 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      841 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      901 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      961 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1021 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1081 tgatttgcgg ccgcaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1141 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1201 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1261 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1321 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1381 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1441 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1501 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1561 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1621 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1681 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1741 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1801 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1861 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1921 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1981 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcaac tatggccatt
     2041 taatgtaaat acttaagaaa aaaaaccaaa ttaattttga tacatgctgc atgtgaaaga
     2101 cccccgctga cgggtagtca atcactcaga ggagaccctc ccaaggaaca gcgagaccac
     2161 aagtcggatg caactgcaag agggtttatt ggatacacgg gtacccgggc gactcagtca
     2221 atcggaggac tggcgccccg agtgaggggt tgtgggctct tttattgagc tcggggagca
     2281 gaagcgcgcg aacagaagcg agaagcgaac tgattggtta gttcaaataa ggcacagggt
     2341 catttcaggt ccttggggca ccctggaaac atctgatggt tctctagaaa ctgctgaggg
     2401 ctggaccgca tctggggacc atctgttctt ggccctgagc cggggcagga actgcttacc
     2461 acagatatcc tgtttggccc atattcagct gttccatctg ttcttggccc tgagccgggg
     2521 caggaactgc ttaccacaga tatcctgttt ggcccatatt cagctgttcc atctgttcct
     2581 gaccttgatc tgaacttctc tattctcagt tatgtatttt tccatgcctt gcaaaatggc
     2641 gttacttaag ctagcttgcc aaacctacag gtggggtctt tcattccccc ctttttctgg
     2701 agactaaata aaatctttta ttttatcggc cctctagatg catgctcgag cggccgccag
     2761 tgtgatggat atctgcagaa ttccagcaca ctggcggccg ttactagtgg atcctgaacc
     2821 cccgccactg cctccgcctt tctcggcaga tctcaggctg atctcgccgc ccatccaggg
     2881 cttgatgatg ccgtcctgtt ccagcagcag agcgccctgc ttgtcgatgc cccggctgat
     2941 gccgaagatc tctttgtcgc cgatgatcag tttcacgggt ctgttgatga agttgtccag
     3001 cttctcccat ctggacaggt agggggccag gccttcctgc tcgaacagtt ccagggcggc
     3061 tctcagctct ctgatcagca tggcggccag ggtgtttctg tccaggttga tgccggcttc
     3121 ctgcagggtg atccagccct gattcacgac ggactcttcc acccgccgca tggccatatt
     3181 gatgccagcg ccaatcacga tctgggcagc gtcgcctgtt ttgcctgtca gttccaccag
     3241 gatgccggcc agcttccggt cctgcagata caggtcgttg ggccacttca cgcgcacttt
     3301 gtcggcgccc agctttctca gcacctcggc catcacgatg ccgatcacga ggctcagtcc
     3361 aatagcggcg gcaggtccct gttccagccg ccagaacatg ctcaggtaca gattggcgcc
     3421 gaagggagag aaccactttc tgcccctgcc gcctctgcca gcctgctgat actcggcgat
     3481 acaggcgtcg ccgcttttca gctcgccgat ccggtccagc aggtactggt tggtgctgtc
     3541 gatcacgggc agcacagcca cagatccgcc atccagctgg cccaggatct gcttggcgtt
     3601 cagcagctgg ataggctcgg gcaggctgta gcccttgcca ggcacggtga acacgtccac
     3661 gccccagtct ctcagggtct ggatgtgctt gttgatggcg gctctagaca tgcccagtgt
     3721 ctcgcccagc tgctcgccag agtggaactc gccattggcc agcagggcga tcagcttcag
     3781 gggcacggtg ttgtccttgt ggtgatggtg atgatgggat cctctctttc ttaacaaatc
     3841 ttcttcagaa atcaactttt gttccatggt ggctgatccg agctcggtac caagcttggg
     3901 tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta
     3961 gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     4021 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     4081 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     4141 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     4201 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     4261 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     4321 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     4381 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     4441 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     4501 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
     4561 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
     4621 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
     4681 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
     4741 tactgagagt gcaccatagg ggatcgggag atccgatgca tggggtcgtg cgctcctttc
     4801 ggtcgggcgc tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg tcatgcacca
     4861 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     4921 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     4981 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     5041 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     5101 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     5161 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     5221 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     5281 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     5341 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     5401 gaggcgcacc gtgggcttgt actcggtcat ggtaagctga tccggccggc gcctagagaa
     5461 ggagtgaggg ctggataaag ggaggatcga ggcggggtcg aacgaggagg ttcaaggggg
     5521 agagacgggg cggatggagg aagaggaggc ggaggcttag ggtgtacaaa gggcttgacc
     5581 cagggagggg ggtcaaaagc caaggcttcc caggtcacga tgtaggggac ctggtctggg
     5641 tgtccatgcg ggccaggtga aaagaccttg atcttaacct gggtgatgag gtctcggtta
     5701 aaggtgccgt ctcgcggcca tccgacgtta aaggttggcc attctgcaga gcagaaggta
     5761 acccaacgtc tcttcttgac atctaccgac tggttgtgag cgatccgctc gacatctttc
     5821 cagtgaccta aggtcaaact taagggagtg gtaacagtct ggccctaatt ttcagacaaa
     5881 tacagaaaca cagtcagaca gagacaacac agaacgatgc tgcagcgctg cagcagacaa
     5941 gacgcgcggc ttcggttcca aaccgaaagc aaaaattcag acggaggcgg gaactgtttt
     6001 aggttctcgt ctcctaccag aaccacatat cctgacgggg tcggattcca catcgactcc
     6061 cttcctcagg tcgggccaca aaaacggccc ccaaagtccc tgggacgtct cccagggttg
     6121 cggccgggtg ttcagaactc gtcagttcca ccacgggtcc gccagataca gagctagtta
     6181 gctaactagt accgacgcag gcgcataaaa tcagtcatag acactagaca atcggacaga
     6241 cacagataag ttgctggcca gcttgcctcc cggtggtggg tcggtggtcc ctgggcaggg
     6301 gtctcccgat cccggacgag cccccaaatg aaagaccccc gctgacgggt agtcaatcac
     6361 tcagaggaga ccctcccaag gaacagcgag accacaagtc ggatgcaact gcaagagggt
     6421 ttattggata cacgggtacc cgggcgactc agtcaatcgg aggactggcg ccccgagtga
     6481 ggggttgtgg gctcttttat tgagctcggg gagcagaagc gcgcgaacag aagcgagaag
     6541 cgaactgatt ggttagttca aataaggcac agggtcattt caggtccttg gggcaccctg
     6601 gaaacatctg atggttctct agaaactgct gagggctgga ccgcatctgg ggaccatctg
     6661 ttcttggccc tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt
     6721 cagctgttcc atctgttctt ggccctgag
//
