LOCUS       sequence-238213-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stably expresses myc-BirA* fused to NRASG12V 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*NRASG12V
FEATURES             Location/Qualifiers
     source          1..6706
                     /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"
     regulatory      1404..1413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1413..1442
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1446..1793,1794..1796,1797..2405)
                     /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"
     primer_bind     complement(3029..3049)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3034..3363
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3214..3349
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3373..3972
                     /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(3373..3392)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3869..3889
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4193..4662
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4792..4809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4963..5551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5043..5062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5711..6502,6503..6571))
                     /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     6334..6353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6572..6676)
                     /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 ccagtgtggt ggtacgtagg
     1381 aattcggaga cccaagctgg ctagccacca tggaacaaaa actcatctca gaagaggatc
     1441 tcgacaagga caacaccgtg cccctgaagc tgatcgccct gctggccaac ggcgagttcc
     1501 actctggcga gcagctggga gagaccctgg gaatgagcag agccgccatc aacaagcaca
     1561 tccagacact gagagactgg ggagtggacg tgttcaccgt gcctggcaag ggctacagcc
     1621 tgcctgagcc tatccagctg ctgaacgcca agcagatcct gggacagctg gatggcggaa
     1681 gcgtggccgt gctgcctgtg atcgactcca ccaatcagta cctgctggac agaatcggag
     1741 agctgaagtc cggcgacgcc tgcatcgccg agtaccagca ggctggcaga ggaggcagag
     1801 gacggaagtg gttcagccca ttcggagcca acctgtacct gtccatgttc tggagactgg
     1861 agcagggacc tgctgctgcc atcggactga gtctggtgat cggaatcgtg atggccgagg
     1921 tgctgagaaa gctgggagcc gacaaggtga gagtgaagtg gcctaatgac ctgtacctcc
     1981 aggaccgcaa gctggctggc atcctggtgg agctgacagg caagacaggc gatgccgctc
     2041 agatcgtgat cggagccgga atcaacatgg ccatgagaag agtggaggag agcgtggtga
     2101 accagggctg gatcaccctg caggaggctg gcatcaacct ggaccggaac accctggccg
     2161 ccatgctgat cagagagctg agagccgctc tggagctgtt cgagcaggag ggactggctc
     2221 cttacctgag cagatgggag aagctggaca acttcatcaa cagacctgtg aagctgatca
     2281 tcggcgacaa ggaaatcttc ggcatctcca gaggaatcga caagcaggga gctctgctgc
     2341 tggagcagga cggaatcatc aagccctgga tgggcggaga aatctccctg agaagcgcag
     2401 agaagctcga gctcgagcgg ccgccactgt gctggatatc tgcagaattc actgagtaca
     2461 aactggtggt ggttggagca gtgggtgttg ggaaaagcgc actgacaatc cagctaatcc
     2521 agaaccactt tgtagatgaa tatgatccca ccatagagga ttcttacaga aaacaagtgg
     2581 ttatagatgg tgaaacctgt ttgttggaca tactggatac agctggacaa gaagagtaca
     2641 gtgccatgag agaccaatac atgaggacag gcgaaggctt cctctgtgta tttgccatca
     2701 ataatagcaa gtcatttgcg gatattaacc tctacaggga gcagattaag cgagtaaaag
     2761 actcggatga tgtacctatg gtgctagtgg gaaacaagtg tgatttgcca acaaggacag
     2821 ttgatacaaa acaagcccac gaactggcca agagttacgg gattccattc attgaaacct
     2881 cagccaagac cagacagggt gttgaagatg ctttttacac actggtaaga gaaatacgcc
     2941 agtaccgaat gaaaaaactc aacagcagtg atgatgggac tcagggttgt atgggattgc
     3001 catgtgtggt gatgtaagtc gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3061 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3121 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3181 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3241 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3301 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3361 aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3421 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3481 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3541 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3601 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3661 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     3721 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3781 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     3841 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3901 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     3961 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     4021 catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     4081 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     4141 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     4201 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4261 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     4321 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     4381 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     4441 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     4501 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     4561 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     4621 tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa
     4681 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat
     4741 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4801 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4861 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4921 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4981 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5041 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5101 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5161 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5221 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5281 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5341 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5401 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5461 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5521 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5581 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5641 gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat gagtaaactt
     5701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5821 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6001 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcagc
//
