LOCUS       sequence-236175-        6963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 6963)
  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 6963)
  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: pcDNA3.1-myc-BirA*-NRASG12V
FEATURES             Location/Qualifiers
     source          1..6963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        156..535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        536..739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     690..710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     784..803
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        784..802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      820..829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             829..858
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(862..1209,1210..1212,1213..1821)
                     /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(2475..2492)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2481..2705
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2751..3179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2838..2857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3048..3069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3188..3208)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3193..3522
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3373..3508
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3435..3454
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3589..4383
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3643..3662)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4253..4272
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4557..4678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4594..4613)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4648..4667
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4727..4743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4727..4743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4740..4762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4751..4767
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4775..4781,4782..4799,4800..4805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4820..4841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4958..4975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5129..5717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5209..5228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5888..6679,6680..6748))
                     /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     6511..6530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6749..6853)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6928..6947)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
       61 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
      121 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      541 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      601 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      661 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      721 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      781 aattaatacg actcactata gggagaccca agctggctag ccaccatgga acaaaaactc
      841 atctcagaag aggatctcga caaggacaac accgtgcccc tgaagctgat cgccctgctg
      901 gccaacggcg agttccactc tggcgagcag ctgggagaga ccctgggaat gagcagagcc
      961 gccatcaaca agcacatcca gacactgaga gactggggag tggacgtgtt caccgtgcct
     1021 ggcaagggct acagcctgcc tgagcctatc cagctgctga acgccaagca gatcctggga
     1081 cagctggatg gcggaagcgt ggccgtgctg cctgtgatcg actccaccaa tcagtacctg
     1141 ctggacagaa tcggagagct gaagtccggc gacgcctgca tcgccgagta ccagcaggct
     1201 ggcagaggag gcagaggacg gaagtggttc agcccattcg gagccaacct gtacctgtcc
     1261 atgttctgga gactggagca gggacctgct gctgccatcg gactgagtct ggtgatcgga
     1321 atcgtgatgg ccgaggtgct gagaaagctg ggagccgaca aggtgagagt gaagtggcct
     1381 aatgacctgt acctccagga ccgcaagctg gctggcatcc tggtggagct gacaggcaag
     1441 acaggcgatg ccgctcagat cgtgatcgga gccggaatca acatggccat gagaagagtg
     1501 gaggagagcg tggtgaacca gggctggatc accctgcagg aggctggcat caacctggac
     1561 cggaacaccc tggccgccat gctgatcaga gagctgagag ccgctctgga gctgttcgag
     1621 caggagggac tggctcctta cctgagcaga tgggagaagc tggacaactt catcaacaga
     1681 cctgtgaagc tgatcatcgg cgacaaggaa atcttcggca tctccagagg aatcgacaag
     1741 cagggagctc tgctgctgga gcaggacgga atcatcaagc cctggatggg cggagaaatc
     1801 tccctgagaa gcgcagagaa gctcgagcgg ccgccactgt gctggatatc tgcagaattc
     1861 atgactgagt acaaactggt ggtggttgga gcagtgggtg ttgggaaaag cgcactgaca
     1921 atccagctaa tccagaacca ctttgtagat gaatatgatc ccaccataga ggattcttac
     1981 agaaaacaag tggttataga tggtgaaacc tgtttgttgg acatactgga tacagctgga
     2041 caagaagagt acagtgccat gagagaccaa tacatgagga caggcgaagg cttcctctgt
     2101 gtatttgcca tcaataatag caagtcattt gcggatatta acctctacag ggagcagatt
     2161 aagcgagtaa aagactcgga tgatgtacct atggtgctag tgggaaacaa gtgtgatttg
     2221 ccaacaagga cagttgatac aaaacaagcc cacgaactgg ccaagagtta cgggattcca
     2281 ttcattgaaa cctcagccaa gaccagacag ggtgttgaag atgcttttta cacactggta
     2341 agagaaatac gccagtaccg aatgaaaaaa ctcaacagca gtgatgatgg gactcagggt
     2401 tgtatgggat tgccatgtgt ggtgatgtaa ggatccgagc tcggtaccaa gcttaagttt
     2461 aaaccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2521 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2581 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2641 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2701 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2881 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3121 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3181 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3241 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3421 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3481 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3541 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4381 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4441 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4501 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4561 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4621 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4681 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4741 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4801 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4861 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4921 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4981 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5041 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5101 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5701 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5761 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5821 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5881 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5941 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6001 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6061 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6121 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6181 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6241 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6301 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6361 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6421 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6481 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6541 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6601 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6661 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6721 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6781 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6841 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6901 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6961 gta
//
