LOCUS       sequence-212849-        5858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To fuse your protein of interest to the C-terminus of BioID2 and use
            in proximity-dependent biotin identification (BioID); a small MCS is
            present downstream of myc tag and BioID2 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  AUTHORS   Kim DI, Jensen SC, Noble KA, Kc B, Roux KH, Motamedchaboki K, Roux 
            KJ
  TITLE     An improved smaller biotin ligase for BioID proximity labeling.
  JOURNAL   Mol Biol Cell. 2016 Feb 24. pii: mbc.E15-12-0844.
  PUBMED    26912792
REFERENCE   2  (bases 1 to 5858)
  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: "Mol Biol 
            Cell. 2016 Feb 24. pii: mbc.E15-12-0844."
COMMENT     Sequence Label: mycBioID2-pBABE-puro
FEATURES             Location/Qualifiers
     source          1..5858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             12..481
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    546..745
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             946..1362
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1292..1314
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1330..1346
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1415..1424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1424..1453
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1457..2152
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     primer_bind     complement(2192..2212)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2197..2526
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2377..2512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2536..3135
                     /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(2536..2555)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3032..3052
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3356..3825
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3955..3972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4126..4714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4206..4225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4874..5665,5666..5734))
                     /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     5497..5516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5735..5839)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgcag cctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
       61 ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
      121 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca
      181 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
      241 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
      301 ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg
      361 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
      421 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
      481 atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg
      541 gaggcaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat
      601 tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt
      661 ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg
      721 ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga
      781 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
      841 ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg
      901 tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc
      961 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
     1021 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
     1081 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
     1141 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
     1201 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
     1261 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
     1321 cgatcctccc tttatccagc cctcactcct tctctaggcg ccggccggat cccagtgtgg
     1381 tggtacgtag gaattcggag acccaagctg gctagccacc atggaacaaa aactcatctc
     1441 agaagaggat ctcgacttca agaacctgat ctggctgaag gaggtggaca gcacccagga
     1501 gagactgaag gagtggaacg tgagctacgg caccgccctg gtggccgaca gacagaccaa
     1561 gggcagaggc ggcctgggca gaaagtggct gagccaggag ggcggcctgt acttcagctt
     1621 cctgctgaac cccaaggagt tcgagaacct gctgcagctg cccctggtgc tgggcctgag
     1681 cgtgagcgag gccctggagg agatcaccga gatccccttc agcctgaagt ggcccaacga
     1741 cgtgtacttc caggagaaga aggtgagcgg cgtgctgtgc gagctgagca aggacaagct
     1801 gatcgtgggc atcggcatca acgtgaacca gagagagatc cccgaggaga tcaaggacag
     1861 agccaccacc ctgtacgaga tcaccggcaa ggactgggac agaaaggagg tgctgctgaa
     1921 ggtgctgaag agaatcagcg agaacctgaa gaagttcaag gagaagagct tcaaggagtt
     1981 caagggcaag atcgagagca agatgctgta cctgggcgag gaggtgaagc tgctgggcga
     2041 gggcaagatc accggcaagc tggtgggcct gagcgagaag ggcggcgccc tgatcctgac
     2101 cgaggagggc atcaaggaga tcctgagcgg cgagttcagc ctgagaagaa gcctcgagtg
     2161 acttaagttt aaaccgctga gtcgaccctg tggaatgtgt gtcagttagg gtgtggaaag
     2221 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2281 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2341 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2401 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2461 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2521 tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     2581 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     2641 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     2701 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     2761 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     2821 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     2881 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     2941 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     3001 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3061 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     3121 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     3181 ccccatgcat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     3241 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     3301 aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa
     3361 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     3421 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     3481 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
     3541 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     3601 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     3661 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     3721 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     3781 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc
     3841 aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg
     3901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4801 aaggatcttc acctagatcc ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa
     4861 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4921 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4981 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5041 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5101 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5161 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5221 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5281 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5341 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5401 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5461 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5521 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5581 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5641 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5701 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5761 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5821 aacaaatagg ggttccgcgc acatttcccc gaaaagtg
//
