LOCUS       sequence-238347-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stably expresses myc-BirA* fused to
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  AUTHORS   Adhikari H, Counter CM
  TITLE     Interrogating the protein interactomes of RAS isoforms identifies 
            PIP5K1A as a KRAS-specific vulnerability.
  JOURNAL   Nat Commun. 2018 Sep 7;9(1):3646. doi: 10.1038/s41467-018-05692-6.
  PUBMED    30194290
REFERENCE   2  (bases 1 to 6706)
  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: "Nat 
            Commun."; date: "2018-09-7"; volume: "9(1):3646. doi"; pages: " 
            10.1038/s41467-018-05692-6"
COMMENT     Sequence Label: mycBioID-pBabePuro-myc-BirA*HRASG12V
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             6..475
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    540..739
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             940..1356
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1286..1308
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1324..1340
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1409..1418
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1418..1447
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1451..1798,1799..1801,1802..2410)
                     /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             join(2456..2485,2486..2488,2489..3022)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase H-Ras"
                     /label=H-Ras (G12V)
                     /translation="TEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVI
                     DGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVKD
                     SDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREIR
                     QHKLRKLNPPDESGPGCMSCKCVLS"
     primer_bind     complement(3034..3054)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3039..3368
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3219..3354
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3378..3977
                     /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(3378..3397)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3874..3894
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4198..4667
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4797..4814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4968..5556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5048..5067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5716..6507,6508..6576))
                     /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     6339..6358
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6577..6681)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
       61 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
      121 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
      181 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
      241 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
      301 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
      361 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
      421 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
      481 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca
      541 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
      601 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
      661 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt
      721 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg
      781 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
      841 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt
      901 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc
      961 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1021 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1081 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1141 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     1201 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1261 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1321 tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcccagt gtggtggtac
     1381 gtaggaattc ggagacccaa gctggctagc caccatggaa caaaaactca tctcagaaga
     1441 ggatctcgac aaggacaaca ccgtgcccct gaagctgatc gccctgctgg ccaacggcga
     1501 gttccactct ggcgagcagc tgggagagac cctgggaatg agcagagccg ccatcaacaa
     1561 gcacatccag acactgagag actggggagt ggacgtgttc accgtgcctg gcaagggcta
     1621 cagcctgcct gagcctatcc agctgctgaa cgccaagcag atcctgggac agctggatgg
     1681 cggaagcgtg gccgtgctgc ctgtgatcga ctccaccaat cagtacctgc tggacagaat
     1741 cggagagctg aagtccggcg acgcctgcat cgccgagtac cagcaggctg gcagaggagg
     1801 cagaggacgg aagtggttca gcccattcgg agccaacctg tacctgtcca tgttctggag
     1861 actggagcag ggacctgctg ctgccatcgg actgagtctg gtgatcggaa tcgtgatggc
     1921 cgaggtgctg agaaagctgg gagccgacaa ggtgagagtg aagtggccta atgacctgta
     1981 cctccaggac cgcaagctgg ctggcatcct ggtggagctg acaggcaaga caggcgatgc
     2041 cgctcagatc gtgatcggag ccggaatcaa catggccatg agaagagtgg aggagagcgt
     2101 ggtgaaccag ggctggatca ccctgcagga ggctggcatc aacctggacc ggaacaccct
     2161 ggccgccatg ctgatcagag agctgagagc cgctctggag ctgttcgagc aggagggact
     2221 ggctccttac ctgagcagat gggagaagct ggacaacttc atcaacagac ctgtgaagct
     2281 gatcatcggc gacaaggaaa tcttcggcat ctccagagga atcgacaagc agggagctct
     2341 gctgctggag caggacggaa tcatcaagcc ctggatgggc ggagaaatct ccctgagaag
     2401 cgcagagaag ctcgagctcg agcggccgcc actgtgctgg atatctgcag aattcacgga
     2461 atataagctg gtggtggtgg gcgccgtggg tgtgggcaag agtgcgctga ccatccagct
     2521 gatccagaac cattttgtgg acgaatacga ccccactata gaggattcct accggaagca
     2581 ggtggtcatt gatggggaga cgtgcctgtt ggacatcctg gataccgccg gccaggagga
     2641 gtacagcgcc atgcgggacc agtacatgcg caccggggag ggcttcctgt gtgtgtttgc
     2701 catcaacaac accaagtctt ttgaggacat ccaccagtac agggagcaga tcaaacgggt
     2761 gaaggactcg gatgacgtgc ccatggtgct ggtggggaac aagtgtgacc tggctgcacg
     2821 cactgtggaa tctcggcagg ctcaggacct cgcccgaagc tacggcatcc cctacatcga
     2881 gacctcggcc aagacccggc agggagtgga ggatgccttc tacacgttgg tgcgtgagat
     2941 ccggcagcac aagctgcgga agctgaaccc tcctgatgag agtggccccg gctgcatgag
     3001 ctgcaagtgt gtgctctcct gagtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa
     3061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3121 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3181 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3241 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3301 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3361 tttgcaaaaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     3421 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     3481 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     3541 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     3601 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     3661 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     3721 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     3781 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     3841 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     3901 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     3961 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     4021 gaccccatgc atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     4081 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     4141 aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg
     4201 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     4261 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     4321 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     4381 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     4441 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     4501 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     4561 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     4621 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta
     4681 tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa
     4741 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4801 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4861 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5521 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5581 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5641 aaaaggatct tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta
     5701 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5761 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5821 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5881 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5941 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6001 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6061 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6121 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6181 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6241 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6301 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6361 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6421 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6481 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6541 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6601 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6661 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
