LOCUS       sequence-238205-        6703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stably expresses myc-BirA* fused to KRASG12V in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  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 6703)
  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*KRASG12V
FEATURES             Location/Qualifiers
     source          1..6703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    527..726
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             927..1343
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1273..1295
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1311..1327
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1396..1405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1405..1434
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1438..1785,1786..1788,1789..2397)
                     /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(3018..3038)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3023..3352
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3203..3338
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3362..3961
                     /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(3362..3381)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3858..3878
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4182..4651
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4781..4798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4952..5540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5032..5051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5700..6491,6492..6560))
                     /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     6323..6342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6561..6665)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6696..6703,1..462)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
       61 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
      121 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
      181 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
      241 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
      301 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
      361 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
      421 tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg
      481 atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggcaagc tggccagcaa
      541 cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt
      601 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      661 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      721 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      781 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      841 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      901 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
      961 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1021 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1081 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1141 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1201 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1261 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1321 ccctcactcc ttctctaggc gccggccgga tcccagtgtg gtggtacgta ggaattcgga
     1381 gacccaagct ggctagccac catggaacaa aaactcatct cagaagagga tctcgacaag
     1441 gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca acggcgagtt ccactctggc
     1501 gagcagctgg gagagaccct gggaatgagc agagccgcca tcaacaagca catccagaca
     1561 ctgagagact ggggagtgga cgtgttcacc gtgcctggca agggctacag cctgcctgag
     1621 cctatccagc tgctgaacgc caagcagatc ctgggacagc tggatggcgg aagcgtggcc
     1681 gtgctgcctg tgatcgactc caccaatcag tacctgctgg acagaatcgg agagctgaag
     1741 tccggcgacg cctgcatcgc cgagtaccag caggctggca gaggaggcag aggacggaag
     1801 tggttcagcc cattcggagc caacctgtac ctgtccatgt tctggagact ggagcaggga
     1861 cctgctgctg ccatcggact gagtctggtg atcggaatcg tgatggccga ggtgctgaga
     1921 aagctgggag ccgacaaggt gagagtgaag tggcctaatg acctgtacct ccaggaccgc
     1981 aagctggctg gcatcctggt ggagctgaca ggcaagacag gcgatgccgc tcagatcgtg
     2041 atcggagccg gaatcaacat ggccatgaga agagtggagg agagcgtggt gaaccagggc
     2101 tggatcaccc tgcaggaggc tggcatcaac ctggaccgga acaccctggc cgccatgctg
     2161 atcagagagc tgagagccgc tctggagctg ttcgagcagg agggactggc tccttacctg
     2221 agcagatggg agaagctgga caacttcatc aacagacctg tgaagctgat catcggcgac
     2281 aaggaaatct tcggcatctc cagaggaatc gacaagcagg gagctctgct gctggagcag
     2341 gacggaatca tcaagccctg gatgggcgga gaaatctccc tgagaagcgc agagaagctc
     2401 gagctcgagc ggccgccact gtgctggata tctgcagaat tcactgaata taaactagtg
     2461 gtggtgggcg ccgtgggcgt gggcaagagc gccctgacca tccagctgat ccagaaccac
     2521 ttcgtggacg agtacgaccc caccatcgag gacagctaca gaaagcaggt ggtgatcgac
     2581 ggcgagacct gcctgctgga catcctggac accgccggcc aggaggagta cagcgccatg
     2641 agagaccagt acatgagaac cggcgagggc ttcctgtgcg tgttcgccat caacaacacc
     2701 aagagcttcg aggacatcca ccactacaga gagcagatca agagagtgaa ggacagcgag
     2761 gacgtgccta tggtgctggt gggcaacaag tgcgacctgc ccagcagaac cgtggacacc
     2821 aagcaggccc aggacctggc cagaagctac ggcatcccct tcatcgagac cagcgccaag
     2881 accagacagg gcgtggacga cgccttctac accctggtga gagagatcag aaagcacaag
     2941 gagaagatga gcaaggacgg caagaagaag aagaagaaga gcaagaccaa gtgcgtgatc
     3001 atgtaagtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3181 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3241 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3301 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     3361 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3421 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     3481 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     3541 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     3601 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     3661 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     3721 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     3781 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     3841 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     3901 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     3961 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     4021 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     4081 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     4141 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     4201 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     4261 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     4321 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     4381 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     4441 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     4501 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     4561 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     4621 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
     4681 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     4741 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5461 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5641 agatcctttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5701 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5761 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5821 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5881 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5941 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6001 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6061 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6121 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6181 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6241 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6301 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6361 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6421 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6481 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6541 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6601 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6661 ccgcgcacat ttccccgaaa agtgccacct gcagcctgaa tat
//
